Beam-induced motion correction for sub-megadalton cryo-EM particles.

Science.gov (United States)

Scheres, Sjors Hw

2014-08-13

In electron cryo-microscopy (cryo-EM), the electron beam that is used for imaging also causes the sample to move. This motion blurs the images and limits the resolution attainable by single-particle analysis. In a previous Research article (Bai et al., 2013) we showed that correcting for this motion by processing movies from fast direct-electron detectors allowed structure determination to near-atomic resolution from 35,000 ribosome particles. In this Research advance article, we show that an improved movie processing algorithm is applicable to a much wider range of specimens. The new algorithm estimates straight movement tracks by considering multiple particles that are close to each other in the field of view, and models the fall-off of high-resolution information content by radiation damage in a dose-dependent manner. Application of the new algorithm to four data sets illustrates its potential for significantly improving cryo-EM structures, even for particles that are smaller than 200 kDa. Copyright © 2014, Scheres.

Correlative cryo-fluorescence light microscopy and cryo-electron tomography of Streptomyces.

Science.gov (United States)

Koning, Roman I; Celler, Katherine; Willemse, Joost; Bos, Erik; van Wezel, Gilles P; Koster, Abraham J

2014-01-01

Light microscopy and electron microscopy are complementary techniques that in a correlative approach enable identification and targeting of fluorescently labeled structures in situ for three-dimensional imaging at nanometer resolution. Correlative imaging allows electron microscopic images to be positioned in a broader temporal and spatial context. We employed cryo-correlative light and electron microscopy (cryo-CLEM), combining cryo-fluorescence light microscopy and cryo-electron tomography, on vitrified Streptomyces bacteria to study cell division. Streptomycetes are mycelial bacteria that grow as long hyphae and reproduce via sporulation. On solid media, Streptomyces subsequently form distinct aerial mycelia where cell division leads to the formation of unigenomic spores which separate and disperse to form new colonies. In liquid media, only vegetative hyphae are present divided by noncell separating crosswalls. Their multicellular life style makes them exciting model systems for the study of bacterial development and cell division. Complex intracellular structures have been visualized with transmission electron microscopy. Here, we describe the methods for cryo-CLEM that we applied for studying Streptomyces. These methods include cell growth, fluorescent labeling, cryo-fixation by vitrification, cryo-light microscopy using a Linkam cryo-stage, image overlay and relocation, cryo-electron tomography using a Titan Krios, and tomographic reconstruction. Additionally, methods for segmentation, volume rendering, and visualization of the correlative data are described. © 2014 Elsevier Inc. All rights reserved.

Time-resolved magnetic imaging in an aberration-corrected, energy-filtered photoemission electron microscope

International Nuclear Information System (INIS)

Nickel, F.; Gottlob, D.M.; Krug, I.P.; Doganay, H.; Cramm, S.; Kaiser, A.M.; Lin, G.; Makarov, D.; Schmidt, O.G.

2013-01-01

We report on the implementation and usage of a synchrotron-based time-resolving operation mode in an aberration-corrected, energy-filtered photoemission electron microscope. The setup consists of a new type of sample holder, which enables fast magnetization reversal of the sample by sub-ns pulses of up to 10 mT. Within the sample holder current pulses are generated by a fast avalanche photo diode and transformed into magnetic fields by means of a microstrip line. For more efficient use of the synchrotron time structure, we developed an electrostatic deflection gating mechanism capable of beam blanking within a few nanoseconds. This allows us to operate the setup in the hybrid bunch mode of the storage ring facility, selecting one or several bright singular light pulses which are temporally well-separated from the normal high-intensity multibunch pulse pattern. - Highlights: • A new time-resolving operation mode in photoemission electron microscopy is shown. • Our setup works within an energy-filtered, aberration-corrected PEEM. • A new gating system for bunch selection using synchrotron radiation is developed. • An alternative magnetic excitation system is developed. • First tr-imaging using an energy-filtered, aberration-corrected PEEM is shown

Design and commissioning of an aberration-corrected ultrafast spin-polarized low energy electron microscope with multiple electron sources.

Science.gov (United States)

Wan, Weishi; Yu, Lei; Zhu, Lin; Yang, Xiaodong; Wei, Zheng; Liu, Jefferson Zhe; Feng, Jun; Kunze, Kai; Schaff, Oliver; Tromp, Ruud; Tang, Wen-Xin

2017-03-01

We describe the design and commissioning of a novel aberration-corrected low energy electron microscope (AC-LEEM). A third magnetic prism array (MPA) is added to the standard AC-LEEM with two prism arrays, allowing the incorporation of an ultrafast spin-polarized electron source alongside the standard cold field emission electron source, without degrading spatial resolution. The high degree of symmetries of the AC-LEEM are utilized while we design the electron optics of the ultrafast spin-polarized electron source, so as to minimize the deleterious effect of time broadening, while maintaining full control of electron spin. A spatial resolution of 2nm and temporal resolution of 10ps (ps) are expected in the future time resolved aberration-corrected spin-polarized LEEM (TR-AC-SPLEEM). The commissioning of the three-prism AC-LEEM has been successfully finished with the cold field emission source, with a spatial resolution below 2nm. Copyright © 2017 Elsevier B.V. All rights reserved.

High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers

International Nuclear Information System (INIS)

Schellenberger, Pascale; Kaufmann, Rainer; Siebert, C. Alistair; Hagen, Christoph; Wodrich, Harald; Grünewald, Kay

2014-01-01

Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell. - Highlights: • Vitrified mammalian cell were imaged by fluorescence and electron cryo microscopy. • TetraSpeck fluorescence markers were added to correct shifts between cryo fluorescence channels. • FluoSpheres fiducials were used as reference points to assign new coordinates to cryoEM images. • Adenovirus particles were localised with an average correlation precision of 63 nm

High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers

Energy Technology Data Exchange (ETDEWEB)

Schellenberger, Pascale [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Kaufmann, Rainer [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU (United Kingdom); Siebert, C. Alistair; Hagen, Christoph [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Wodrich, Harald [Microbiologie Fondamentale et Pathogénicité, MFP CNRS UMR 5234, University of Bordeaux SEGALEN, 146 rue Leo Seignat, 33076 Bordeaux (France); Grünewald, Kay, E-mail: kay@strubi.ox.ac.uk [Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)

2014-08-01

Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell. - Highlights: • Vitrified mammalian cell were imaged by fluorescence and electron cryo microscopy. • TetraSpeck fluorescence markers were added to correct shifts between cryo fluorescence channels. • FluoSpheres fiducials were used as reference points to assign new coordinates to cryoEM images. • Adenovirus particles were localised with an average correlation precision of 63 nm.

Aberration-corrected STEM: current performance and future directions

Energy Technology Data Exchange (ETDEWEB)

Nellist, P D [Department of Physics, University of Dublin, Trinity College, Dublin 2 (Ireland); Chisholm, M F [Condensed Matter Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6030 (United States); Lupini, A R [Condensed Matter Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6030 (United States); Borisevich, A [Condensed Matter Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6030 (United States); Jr, W H Sides [Condensed Matter Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6030 (United States); Pennycook, S J [Condensed Matter Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6030 (United States); Dellby, N [Nion Co., 1102 8th St., Kirkland, WA 98033 (United States); Keyse, R [Nion Co., 1102 8th St., Kirkland, WA 98033 (United States); Krivanek, O L [Nion Co., 1102 8th St., Kirkland, WA 98033 (United States); Murfitt, M F [Nion Co., 1102 8th St., Kirkland, WA 98033 (United States); Szilagyi, Z S [Nion Co., 1102 8th St., Kirkland, WA 98033 (United States)

2006-02-22

Through the correction of spherical aberration in the scanning transmission electron microscope (STEM), the resolving of a 78 pm atomic column spacing has been demonstrated along with information transfer to 61 pm. The achievement of this resolution required careful control of microscope instabilities, parasitic aberrations and the compensation of uncorrected, higher order aberrations. Many of these issues are improved in a next generation STEM fitted with a new design of aberration corrector, and an initial result demonstrating aberration correction to a convergence semi-angle of 40 mrad is shown. The improved spatial resolution and beam convergence allowed for by such correction has implications for the way in which experiments are performed and how STEM data should be interpreted.

Correlated cryo-fluorescence and cryo-electron microscopy with high spatial precision and improved sensitivity

International Nuclear Information System (INIS)

Schorb, Martin; Briggs, John A.G.

2014-01-01

Performing fluorescence microscopy and electron microscopy on the same sample allows fluorescent signals to be used to identify and locate features of interest for subsequent imaging by electron microscopy. To carry out such correlative microscopy on vitrified samples appropriate for structural cryo-electron microscopy it is necessary to perform fluorescence microscopy at liquid-nitrogen temperatures. Here we describe an adaptation of a cryo-light microscopy stage to permit use of high-numerical aperture objectives. This allows high-sensitivity and high-resolution fluorescence microscopy of vitrified samples. We describe and apply a correlative cryo-fluorescence and cryo-electron microscopy workflow together with a fiducial bead-based image correlation procedure. This procedure allows us to locate fluorescent bacteriophages in cryo-electron microscopy images with an accuracy on the order of 50 nm, based on their fluorescent signal. It will allow the user to precisely and unambiguously identify and locate objects and events for subsequent high-resolution structural study, based on fluorescent signals. - Highlights: • Workflow for correlated cryo-fluorescence and cryo-electron microscopy. • Cryo-fluorescence microscopy setup incorporating a high numerical aperture objective. • Fluorescent signals located in cryo-electron micrographs with 50 nm spatial precision

Correlated cryo-fluorescence and cryo-electron microscopy with high spatial precision and improved sensitivity

Energy Technology Data Exchange (ETDEWEB)

Schorb, Martin [Structural and Computational Biology Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany); Briggs, John A.G., E-mail: john.briggs@embl.de [Structural and Computational Biology Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany); Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany)

2014-08-01

Performing fluorescence microscopy and electron microscopy on the same sample allows fluorescent signals to be used to identify and locate features of interest for subsequent imaging by electron microscopy. To carry out such correlative microscopy on vitrified samples appropriate for structural cryo-electron microscopy it is necessary to perform fluorescence microscopy at liquid-nitrogen temperatures. Here we describe an adaptation of a cryo-light microscopy stage to permit use of high-numerical aperture objectives. This allows high-sensitivity and high-resolution fluorescence microscopy of vitrified samples. We describe and apply a correlative cryo-fluorescence and cryo-electron microscopy workflow together with a fiducial bead-based image correlation procedure. This procedure allows us to locate fluorescent bacteriophages in cryo-electron microscopy images with an accuracy on the order of 50 nm, based on their fluorescent signal. It will allow the user to precisely and unambiguously identify and locate objects and events for subsequent high-resolution structural study, based on fluorescent signals. - Highlights: • Workflow for correlated cryo-fluorescence and cryo-electron microscopy. • Cryo-fluorescence microscopy setup incorporating a high numerical aperture objective. • Fluorescent signals located in cryo-electron micrographs with 50 nm spatial precision.

Third-rank chromatic aberrations of electron lenses.

Science.gov (United States)

Liu, Zhixiong

2018-02-01

In this paper the third-rank chromatic aberration coefficients of round electron lenses are analytically derived and numerically calculated by Mathematica. Furthermore, the numerical results are cross-checked by the differential algebraic (DA) method, which verifies that all the formulas for the third-rank chromatic aberration coefficients are completely correct. It is hoped that this work would be helpful for further chromatic aberration correction in electron microscopy. Copyright © 2017 Elsevier B.V. All rights reserved.

A new aberration-corrected, energy-filtered LEEM/PEEM instrument. I. Principles and design

International Nuclear Information System (INIS)

Tromp, R.M.; Hannon, J.B.; Ellis, A.W.; Wan, W.; Berghaus, A.; Schaff, O.

2010-01-01

We describe a new design for an aberration-corrected low energy electron microscope (LEEM) and photo electron emission microscope (PEEM), equipped with an in-line electron energy filter. The chromatic and spherical aberrations of the objective lens are corrected with an electrostatic electron mirror that provides independent control over the chromatic and spherical aberration coefficients C c and C 3 , as well as the mirror focal length, to match and correct the aberrations of the objective lens. For LEEM (PEEM) the theoretical resolution is calculated to be ∼1.5 nm (∼4 nm). Unlike previous designs, this instrument makes use of two magnetic prism arrays to guide the electron beam from the sample to the electron mirror, removing chromatic dispersion in front of the mirror by symmetry. The aberration correction optics was retrofitted to an uncorrected instrument with a base resolution of 4.1 nm in LEEM. Initial results in LEEM show an improvement in resolution to ∼2 nm.

Brief history of the Cambridge STEM aberration correction project and its progeny

Energy Technology Data Exchange (ETDEWEB)

Brown, L. Michael [Cavendish Laboratory, University of Cambridge, J.J. Thomson Avenue, Cambridge CB3 0HE (United Kingdom); Batson, Philip E. [Institute for Advanced Materials, Devices and Nanotechnology, Rutgers University, Piscataway, NJ 08854 (United States); Department of Physics, Rutgers University, Piscataway, NJ 08854 (United States); Department of Materials Science, Rutgers University, Piscataway, NJ 08854 (United States); Dellby, Niklas [Nion Company, 11515 NE 118th Street, Kirkland, WA 98034 (United States); Krivanek, Ondrej L. [Nion Company, 11515 NE 118th Street, Kirkland, WA 98034 (United States); Department of Physics, Arizona State University, Tempe, AZ 85287 (United States)

2015-10-15

We provide a brief history of the project to correct the spherical aberration of the scanning transmission electron microscope (STEM) that started in Cambridge (UK) and continued in Kirkland (WA, USA), Yorktown Heights (NY, USA), and other places. We describe the project in the full context of other aberration correction research and related work, partly in response to the incomplete context presented in the paper “In quest of perfection in electron optics: A biographical sketch of Harald Rose on the occasion of his 80th birthday”, recently published in Ultramicroscopy. - Highlights: • We provide a brief history of the Cambridge project to correct the spherical aberration of the scanning transmission electron microscope (STEM). • We describe the project in the full context of other aberration correction work and related research. • We summarize our corrector development work that followed the Cambridge project, and which was the first to reach higher spatial resolution than any non-corrected electron microscope.

Brief history of the Cambridge STEM aberration correction project and its progeny

International Nuclear Information System (INIS)

Brown, L. Michael; Batson, Philip E.; Dellby, Niklas; Krivanek, Ondrej L.

2015-01-01

We provide a brief history of the project to correct the spherical aberration of the scanning transmission electron microscope (STEM) that started in Cambridge (UK) and continued in Kirkland (WA, USA), Yorktown Heights (NY, USA), and other places. We describe the project in the full context of other aberration correction research and related work, partly in response to the incomplete context presented in the paper “In quest of perfection in electron optics: A biographical sketch of Harald Rose on the occasion of his 80th birthday”, recently published in Ultramicroscopy. - Highlights: • We provide a brief history of the Cambridge project to correct the spherical aberration of the scanning transmission electron microscope (STEM). • We describe the project in the full context of other aberration correction work and related research. • We summarize our corrector development work that followed the Cambridge project, and which was the first to reach higher spatial resolution than any non-corrected electron microscope

High-resolution TEM and the application of direct and indirect aberration correction

DEFF Research Database (Denmark)

Hetherington, Crispin J.D.; Chang, Lan-Yun Shery; Haigh, Sarah

2008-01-01

Aberration correction leads to a substantial improvement in the directly interpretable resolution of transmission electron microscopes. Correction of the aberrations has been achieved electron-optically through a hexapole-based corrector and also indirectly by computational analysis of a focal or...

The objective lens of the electron microscope with correction of spherical and axial chromatic aberrations.

Science.gov (United States)

Bimurzaev, S B; Aldiyarov, N U; Yakushev, E M

2017-10-01

The paper describes the principle of operation of a relatively simple aberration corrector for the transmission electron microscope objective lens. The electron-optical system of the aberration corrector consists of the two main elements: an electrostatic mirror with rotational symmetry and a magnetic deflector formed by the round-shaped magnetic poles. The corrector operation is demonstrated by calculations on the example of correction of basic aberrations of the well-known objective lens with a bell-shaped distribution of the axial magnetic field. Two of the simplest versions of the corrector are considered: a corrector with a two-electrode electrostatic mirror and a corrector with a three-electrode electrostatic mirror. It is shown that using the two-electrode mirror one can eliminate either spherical or chromatic aberration of the objective lens, without changing the value of its linear magnification. Using a three-electrode mirror, it is possible to eliminate spherical and chromatic aberrations of the objective lens simultaneously, which is especially important in designing electron microscopes with extremely high resolution. © The Author 2017. 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.

Progress toward an aberration-corrected low energy electron microscope for DNA sequencing and surface analysis.

Science.gov (United States)

Mankos, Marian; Shadman, Khashayar; N'diaye, Alpha T; Schmid, Andreas K; Persson, Henrik H J; Davis, Ronald W

2012-11-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel imaging technique aimed at high resolution imaging of macromolecules, nanoparticles, and surfaces. MAD-LEEM combines three innovative electron-optical concepts in a single tool: a monochromator, a mirror aberration corrector, and dual electron beam illumination. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. The aberration corrector is needed to achieve subnanometer resolution at landing energies of a few hundred electronvolts. The dual flood illumination approach eliminates charging effects generated when a conventional, single-beam LEEM is used to image insulating specimens. The low landing energy of electrons in the range of 0 to a few hundred electronvolts is also critical for avoiding radiation damage, as high energy electrons with kilo-electron-volt kinetic energies cause irreversible damage to many specimens, in particular biological molecules. The performance of the key electron-optical components of MAD-LEEM, the aberration corrector combined with the objective lens and a magnetic beam separator, was simulated. Initial results indicate that an electrostatic electron mirror has negative spherical and chromatic aberration coefficients that can be tuned over a large parameter range. The negative aberrations generated by the electron mirror can be used to compensate the aberrations of the LEEM objective lens for a range of electron energies and provide a path to achieving subnanometer spatial resolution. First experimental results on characterizing DNA molecules immobilized on Au substrates in a LEEM are presented. Images obtained in a spin-polarized LEEM demonstrate that high contrast is achievable at low electron energies in the range of 1-10 eV and show that small changes in landing energy have a strong impact on the achievable contrast. The MAD-LEEM approach

Determination of aberration center of Ronchigram for automated aberration correctors in scanning transmission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Sannomiya, Takumi, E-mail: sannomiya@mtl.titech.ac.jp [Tokyo Institute of Technology, Ookayama, Tokyo (Japan); Sawada, Hidetaka; Nakamichi, Tomohiro; Hosokawa, Fumio [JEOL Limited, Akishima, Tokyo (Japan); Nakamura, Yoshio; Tanishiro, Yasumasa; Takayanagi, Kunio [Tokyo Institute of Technology, Ookayama, Tokyo (Japan)

2013-12-15

A generic method to determine the aberration center is established, which can be utilized for aberration calculation and axis alignment for aberration corrected electron microscopes. In this method, decentering induced secondary aberrations from inherent primary aberrations are minimized to find the appropriate axis center. The fitness function to find the optimal decentering vector for the axis was defined as a sum of decentering induced secondary aberrations with properly distributed weight values according to the aberration order. Since the appropriate decentering vector is determined from the aberration values calculated at an arbitrary center axis, only one aberration measurement is in principle required to find the center, resulting in /very fast center search. This approach was tested for the Ronchigram based aberration calculation method for aberration corrected scanning transmission electron microscopy. Both in simulation and in experiments, the center search was confirmed to work well although the convergence to find the best axis becomes slower with larger primary aberrations. Such aberration center determination is expected to fully automatize the aberration correction procedures, which used to require pre-alignment of experienced users. This approach is also applicable to automated aperture positioning. - Highlights: • A generic method to determine the aberration center is established for (S)TEM. • Decentering induced secondary aberrations are utilized to find the center. • The method is tested on Ronchigrams both in simulation and experiment. • Proper weighting of the aberration gives a good convergence. • Larger primary aberration results in a slower convergence.

A new aberration-corrected, energy-filtered LEEM/PEEM instrument II. Operation and results

International Nuclear Information System (INIS)

Tromp, R.M.; Hannon, J.B.; Wan, W.; Berghaus, A.; Schaff, O.

2013-01-01

In Part I we described a new design for an aberration-corrected Low Energy Electron Microscope (LEEM) and Photo Electron Emission Microscope (PEEM) equipped with an in-line electron energy filter. The chromatic and spherical aberrations of the objective lens are corrected with an electrostatic electron mirror that provides independent control of the chromatic and spherical aberration coefficients C c and C 3 , as well as the mirror focal length. In this Part II we discuss details of microscope operation, how the microscope is set up in a systematic fashion, and we present typical results. - Highlights: ► The C c and C 3 aberrations of a LEEM/PEEM instrument are corrected with an electrostatic electron mirror. ► The mirror provides independent control over C c , C 3 and focal length in close agreement with theory. ► A detailed alignment procedure for the corrected microscope is given. ► Novel methods to measure C c and C 3 of the objective lens and the mirror are presented. ► We demonstrate a record spatial resolution of 2 nm

High-resolution electron microscopy

CERN Document Server

Spence, John C H

2013-01-01

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

Optimization and stability of the contrast transfer function in aberration-corrected electron microscopy

International Nuclear Information System (INIS)

Tromp, R.M.; Schramm, S.M.

2013-01-01

The Contrast Transfer Function (CTF) describes the manner in which the electron microscope modifies the object exit wave function as a result of objective lens aberrations. For optimum resolution in C 3 -corrected microscopes it is well established that a small negative value of C 3 , offset by positive values of C 5 and defocus C 1 results in the most optimal instrument resolution, and optimization of the CTF has been the subject of several studies. Here we describe a simple design procedure for the CTF that results in a most even transfer of information below the resolution limit. We address not only the resolution of the instrument, but also the stability of the CTF in the presence of small disturbances in C 1 and C 3 . We show that resolution can be traded for stability in a rational and transparent fashion. These topics are discussed quantitatively for both weak-phase and strong-phase (or amplitude) objects. The results apply equally to instruments at high electron energy (TEM) and at very low electron energy (LEEM), as the basic optical properties of the imaging lenses are essentially identical. - Highlights: ► An optimized Contrast Transfer Function for aberration corrected electron microscopes is proposed. ► Based on the properties of the CTF near optimum settings, we address its stability. ► Over some range of parameters resolution can be traded for stability. ► These issues are addressed for weak-phase objects, as well as strong-phase and amplitude object. ► We compare our results with CTF settings previously proposed

Collaborative Computational Project for Electron cryo-Microscopy

Energy Technology Data Exchange (ETDEWEB)

Wood, Chris; Burnley, Tom [Science and Technology Facilities Council, Research Complex at Harwell, Didcot OX11 0FA (United Kingdom); Patwardhan, Ardan [European Molecular Biology Laboratory, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD (United Kingdom); Scheres, Sjors [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom); Topf, Maya [University of London, Malet Street, London WC1E 7HX (United Kingdom); Roseman, Alan [University of Manchester, Oxford Road, Manchester M13 9PT (United Kingdom); Winn, Martyn, E-mail: martyn.winn@stfc.ac.uk [Science and Technology Facilities Council, Daresbury Laboratory, Warrington WA4 4AD (United Kingdom); Science and Technology Facilities Council, Research Complex at Harwell, Didcot OX11 0FA (United Kingdom)

2015-01-01

The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) is a new initiative for the structural biology community, following the success of CCP4 for macromolecular crystallography. Progress in supporting the users and developers of cryoEM software is reported. The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) has recently been established. The aims of the project are threefold: to build a coherent cryoEM community which will provide support for individual scientists and will act as a focal point for liaising with other communities, to support practising scientists in their use of cryoEM software and finally to support software developers in producing and disseminating robust and user-friendly programs. The project is closely modelled on CCP4 for macromolecular crystallography, and areas of common interest such as model fitting, underlying software libraries and tools for building program packages are being exploited. Nevertheless, cryoEM includes a number of techniques covering a large range of resolutions and a distinct project is required. In this article, progress so far is reported and future plans are discussed.

Collaborative Computational Project for Electron cryo-Microscopy

International Nuclear Information System (INIS)

Wood, Chris; Burnley, Tom; Patwardhan, Ardan; Scheres, Sjors; Topf, Maya; Roseman, Alan; Winn, Martyn

2015-01-01

The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) is a new initiative for the structural biology community, following the success of CCP4 for macromolecular crystallography. Progress in supporting the users and developers of cryoEM software is reported. The Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) has recently been established. The aims of the project are threefold: to build a coherent cryoEM community which will provide support for individual scientists and will act as a focal point for liaising with other communities, to support practising scientists in their use of cryoEM software and finally to support software developers in producing and disseminating robust and user-friendly programs. The project is closely modelled on CCP4 for macromolecular crystallography, and areas of common interest such as model fitting, underlying software libraries and tools for building program packages are being exploited. Nevertheless, cryoEM includes a number of techniques covering a large range of resolutions and a distinct project is required. In this article, progress so far is reported and future plans are discussed

Aberration-corrected STEM/TEM imaging at 15 kV

International Nuclear Information System (INIS)

Sasaki, Takeo; Sawada, Hidetaka; Hosokawa, Fumio; Sato, Yuta; Suenaga, Kazu

2014-01-01

The performance of aberration-corrected (scanning) transmission electron microscopy (S/TEM) at an accelerating voltage of 15 kV was evaluated in a low-voltage microscope equipped with a cold-field emission gun and a higher-order aberration corrector. Aberrations up to the fifth order were corrected by the aberration measurement and auto-correction system using the diffractogram tableau method in TEM and Ronchigram analysis in STEM. TEM observation of nanometer-sized particles demonstrated that aberrations up to an angle of 50 mrad were compensated. A TEM image of Si[110] exhibited lattice fringes with a spacing of 0.192 nm, and the power spectrum of the image showed spots corresponding to distances of 0.111 nm. An annular dark-field STEM image of Si[110] showed lattice fringes of (111) and (22¯0) planes corresponding to lattice distances of 0.314 nm and 0.192 nm, respectively. At an accelerating voltage of 15 kV, the developed low-voltage microscope achieved atomic-resolution imaging with a small chromatic aberration and a large uniform phase. - Highlights: • Aberration-corrected STEM/TEM imaging at 15 kV demonstrated lattice fringes of Si[110] single crystal with a spacing of 0.192 nm. • To achieve this performance at a lower accelerating voltage, uniform phase area over 50 mrad is mandatory in Ronchigram and Diffractogram tableau. • This means a higher-order aberration of six-fold astigmatism should be compensated. • In addition, decreasing the effect of chromatic aberration plays an important role for improving the performance of linear scattering component at 15 kV TEM

Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope.

Science.gov (United States)

Wang, Peng; Behan, Gavin; Kirkland, Angus I; Nellist, Peter D; Cosgriff, Eireann C; D'Alfonso, Adrian J; Morgan, Andrew J; Allen, Leslie J; Hashimoto, Ayako; Takeguchi, Masaki; Mitsuishi, Kazutaka; Shimojo, Masayuki

2011-06-01

Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored. Copyright © 2010 Elsevier B.V. All rights reserved.

Electron-beam-induced-current and active secondary-electron voltage-contrast with aberration-corrected electron probes

Energy Technology Data Exchange (ETDEWEB)

Han, Myung-Geun, E-mail: mghan@bnl.gov [Condensed Matter Physics & Materials Science, Brookhaven National Laboratory, Upton, NY 11973 (United States); Garlow, Joseph A. [Condensed Matter Physics & Materials Science, Brookhaven National Laboratory, Upton, NY 11973 (United States); Materials Science and Engineering Department, Stony Brook University, Stony Brook, NY 11794 (United States); Marshall, Matthew S.J.; Tiano, Amanda L. [Department of Chemistry, Stony Brook University, Stony Brook, NY 11974 (United States); Wong, Stanislaus S. [Condensed Matter Physics & Materials Science, Brookhaven National Laboratory, Upton, NY 11973 (United States); Department of Chemistry, Stony Brook University, Stony Brook, NY 11974 (United States); Cheong, Sang-Wook [Department of Physics and Astronomy, Rutgers Center for Emergent Materials, Rutgers University, Piscataway, NJ 08854 (United States); Walker, Frederick J.; Ahn, Charles H. [Department of Applied Physics and Center for Research on Interface Structures and Phenomena, Yale University, New Haven, CT 06520 (United States); Department of Mechanical Engineering and Materials Science, Yale University, New Haven, CT 06520 (United States); Zhu, Yimei [Condensed Matter Physics & Materials Science, Brookhaven National Laboratory, Upton, NY 11973 (United States)

2017-05-15

Highlights: • Electron-beam-induced-current (EBIC) and active secondary-electron voltage-contrast (SE-VC) are demonstrated in STEM mode combined with in situ electrical biasing in a TEM. • Electrostatic potential maps in ferroelectric thin films, multiferroic nanowires, and single crystals obtained by off-axis electron holography were compared with EBIC and SE-VC data. • Simultaneous EBIC and active SE-VC performed with atomic resolution STEM are demonstrated. - Abstract: The ability to map out electrostatic potentials in materials is critical for the development and the design of nanoscale electronic and spintronic devices in modern industry. Electron holography has been an important tool for revealing electric and magnetic field distributions in microelectronics and magnetic-based memory devices, however, its utility is hindered by several practical constraints, such as charging artifacts and limitations in sensitivity and in field of view. In this article, we report electron-beam-induced-current (EBIC) and secondary-electron voltage-contrast (SE-VC) with an aberration-corrected electron probe in a transmission electron microscope (TEM), as complementary techniques to electron holography, to measure electric fields and surface potentials, respectively. These two techniques were applied to ferroelectric thin films, multiferroic nanowires, and single crystals. Electrostatic potential maps obtained by off-axis electron holography were compared with EBIC and SE-VC to show that these techniques can be used as a complementary approach to validate quantitative results obtained from electron holography analysis.

Cryo-electron tomography of bacterial viruses

Energy Technology Data Exchange (ETDEWEB)

Guerrero-Ferreira, Ricardo C. [Division of Pediatric Infectious Diseases, Emory University School of Medicine, Children' s Healthcare of Atlanta, Atlanta, GA 30322 (United States); Wright, Elizabeth R., E-mail: erwrigh@emory.edu [Division of Pediatric Infectious Diseases, Emory University School of Medicine, Children' s Healthcare of Atlanta, Atlanta, GA 30322 (United States)

2013-01-05

Bacteriophage particles contain both simple and complex macromolecular assemblages and machines that enable them to regulate the infection process under diverse environmental conditions with a broad range of bacterial hosts. Recent developments in cryo-electron tomography (cryo-ET) make it possible to observe the interactions of bacteriophages with their host cells under native-state conditions at unprecedented resolution and in three-dimensions. This review describes the application of cryo-ET to studies of bacteriophage attachment, genome ejection, assembly and egress. Current topics of investigation and future directions in the field are also discussed.

Iteration of ultrasound aberration correction methods

Science.gov (United States)

Maasoey, Svein-Erik; Angelsen, Bjoern; Varslot, Trond

2004-05-01

Aberration in ultrasound medical imaging is usually modeled by time-delay and amplitude variations concentrated on the transmitting/receiving array. This filter process is here denoted a TDA filter. The TDA filter is an approximation to the physical aberration process, which occurs over an extended part of the human body wall. Estimation of the TDA filter, and performing correction on transmit and receive, has proven difficult. It has yet to be shown that this method works adequately for severe aberration. Estimation of the TDA filter can be iterated by retransmitting a corrected signal and re-estimate until a convergence criterion is fulfilled (adaptive imaging). Two methods for estimating time-delay and amplitude variations in receive signals from random scatterers have been developed. One method correlates each element signal with a reference signal. The other method use eigenvalue decomposition of the receive cross-spectrum matrix, based upon a receive energy-maximizing criterion. Simulations of iterating aberration correction with a TDA filter have been investigated to study its convergence properties. A weak and strong human-body wall model generated aberration. Both emulated the human abdominal wall. Results after iteration improve aberration correction substantially, and both estimation methods converge, even for the case of strong aberration.

The application of aberration-corrected electron microscopy to the characterization of gold-based catalysts

Science.gov (United States)

Herzing, Andrew A.

Electron microscopy has long been used to study the morphology of heterogeneous catalysts. Recent advances in electron optics now allow for the correction of the inherent spherical aberration (Cs) produced by the objective lens in the scanning transmission electron microscope (STEM, resulting in a significantly improved spatial resolution as well as the ability to use a much larger probe-current than was previously possible. In this thesis, the combination of high-angle annular dark-field (HAADF) imaging and microanalysis by x-ray energy dispersive spectroscopy (XEDS) in an aberration-corrected STEM has been applied for the first time to the characterization of gold-based heterogeneous catalysts. Multi-variate statistical analysis (MSA) has been employed in order to further improve the STEM-XEDS spectrum image data acquired with this technique. In addition, supplemental analysis using electron-energy loss spectroscopy (EELS) and energy-filtered transmission electron microscopy (EFTEM) in an aberration-corrected instrument has also been attempted. These techniques have proven extremely valuable in providing complimentary information to more traditional catalyst characterization techniques such as x-ray photoelectron spectroscopy and x-ray diffraction in four specific problems relating to catalysis. Firstly, the atomic-scale resolution of Cs-corrected HAADF imaging has been utilized to study Au/FeOx catalysts in order to determine the size and structure of the Au clusters present on the support surface. It was discovered that, while both inactive and active catalysts for low-temperature CO oxidation contained large Au particles (> 5 nm) and individual Au atoms, the active catalyst also contained sub-nm clusters comprised of only a few Au atoms. Secondly, novel CeO2 support materials for Au and Au-Pd catalysts were synthesized by precipitation with supercritical CO2. These supports were found to produce significantly more active catalysts than those based on CeO2

Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope

Energy Technology Data Exchange (ETDEWEB)

Wang, Peng; Behan, Gavin; Kirkland, Angus I. [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Nellist, Peter D., E-mail: peter.nellist@materials.ox.ac.uk [Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH (United Kingdom); Cosgriff, Eireann C.; D' Alfonso, Adrian J.; Morgan, Andrew J.; Allen, Leslie J. [School of Physics, University of Melbourne, Parkville, Victoria 3010 (Australia); Hashimoto, Ayako [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Takeguchi, Masaki [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Mitsuishi, Kazutaka [Advanced Nano-characterization Center, National Institute for Materials Science (NIMS), 3-13 Sakura, Tsukuba 305-0003 (Japan); Quantum Dot Research Center, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Shimojo, Masayuki [High Voltage Electron Microscopy Station, NIMS, 3-13 Sakura, Tsukuba 305-0003 (Japan); Advanced Science Research Laboratory, Saitama Institute of Technology, 1690 Fusaiji, Fukaya 369-0293 (Japan)

2011-06-15

Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored. -- Research Highlights: {yields} The confocal probe image in a scanning confocal electron microscopy image reveals information about the thickness and height of the crystalline layer. {yields} The form of the contrast in a three-dimensional bright-field scanning confocal electron microscopy image can be explained in terms of the confocal probe image. {yields} Despite the complicated form of the contrast in bright-field scanning confocal electron microscopy, we see that depth information is transferred on a 10 nm scale.

Advances in cryo-electron tomography for biology and medicine.

Science.gov (United States)

Koning, Roman I; Koster, Abraham J; Sharp, Thomas H

2018-05-01

Cryo-electron tomography (CET) utilizes a combination of specimen cryo-fixation and multi-angle electron microscopy imaging to produce three-dimensional (3D) volume reconstructions of native-state macromolecular and subcellular biological structures with nanometer-scale resolution. In recent years, cryo-electron microscopy (cryoEM) has experienced a dramatic increase in the attainable resolution of 3D reconstructions, resulting from technical improvements of electron microscopes, improved detector sensitivity, the implementation of phase plates, automated data acquisition schemes, and improved image reconstruction software and hardware. These developments also greatly increased the usability and applicability of CET as a diagnostic and research tool, which is now enabling structural biologists to determine the structure of proteins in their native cellular environment to sub-nanometer resolution. These recent technical developments have stimulated us to update on our previous review (Koning, R.I., Koster, A.J., 2009. Cryo-electron tomography in biology and medicine. Ann Anat 191, 427-445) in which we described the fundamentals of CET. In this follow-up, we extend this basic description in order to explain the aforementioned recent advances, and describe related 3D techniques that can be applied to the anatomy of biological systems that are relevant for medicine. Copyright © 2018 Elsevier GmbH. All rights reserved.

Axial geometrical aberration correction up to 5th order with N-SYLC.

Science.gov (United States)

Hoque, Shahedul; Ito, Hiroyuki; Takaoka, Akio; Nishi, Ryuji

2017-11-01

We present N-SYLC (N-fold symmetric line currents) models to correct 5th order axial geometrical aberrations in electron microscopes. In our previous paper, we showed that 3rd order spherical aberration can be corrected by 3-SYLC doublet. After that, mainly the 5th order aberrations remain to limit the resolution. In this paper, we extend the doublet to quadruplet models also including octupole and dodecapole fields for correcting these higher order aberrations, without introducing any new unwanted ones. We prove the validity of our models by analytical calculations. Also by computer simulations, we show that for beam energy of 5keV and initial angle 10mrad at the corrector object plane, beam size of less than 0.5nm is achieved at the corrector image plane. Copyright © 2017 Elsevier B.V. All rights reserved.

Modeling of Image Formation in Cryo-Electron Microscopy

NARCIS (Netherlands)

Vulovic, M.

2013-01-01

Knowledge of the structure of biological specimens is crucial for understanding life. Cryo-electron microscopy (cryo-EM) permits structural studies of biological specimen at their near-native state. The research performed in this thesis represents one of two subprojects of the FOM industrial

Cryo-electron microscopy of membrane proteins.

Science.gov (United States)

Goldie, Kenneth N; Abeyrathne, Priyanka; Kebbel, Fabian; Chami, Mohamed; Ringler, Philippe; Stahlberg, Henning

2014-01-01

Electron crystallography is used to study membrane proteins in the form of planar, two-dimensional (2D) crystals, or other crystalline arrays such as tubular crystals. This method has been used to determine the atomic resolution structures of bacteriorhodopsin, tubulin, aquaporins, and several other membrane proteins. In addition, a large number of membrane protein structures were studied at a slightly lower resolution, whereby at least secondary structure motifs could be identified.In order to conserve the structural details of delicate crystalline arrays, cryo-electron microscopy (cryo-EM) allows imaging and/or electron diffraction of membrane proteins in their close-to-native state within a lipid bilayer membrane.To achieve ultimate high-resolution structural information of 2D crystals, meticulous sample preparation for electron crystallography is of outmost importance. Beam-induced specimen drift and lack of specimen flatness can severely affect the attainable resolution of images for tilted samples. Sample preparations that sandwich the 2D crystals between symmetrical carbon films reduce the beam-induced specimen drift, and the flatness of the preparations can be optimized by the choice of the grid material and the preparation protocol.Data collection in the cryo-electron microscope using either the imaging or the electron diffraction mode has to be performed applying low-dose procedures. Spot-scanning further reduces the effects of beam-induced drift. Data collection using automated acquisition schemes, along with improved and user-friendlier data processing software, is increasingly being used and is likely to bring the technique to a wider user base.

Interaction between single gold atom and the graphene edge: A study via aberration-corrected transmission electron microscopy

KAUST Repository

Wang, Hongtao

2012-01-01

Interaction between single noble metal atoms and graphene edges has been investigated via aberration-corrected and monochromated transmission electron microscopy. A collective motion of the Au atom and the nearby carbon atoms is observed in transition between energy-favorable configurations. Most trapping and detrapping processes are assisted by the dangling carbon atoms, which are more susceptible to knock-on displacements by electron irradiation. Thermal energy is lower than the activation barriers in transition among different energy-favorable configurations, which suggests electron-beam irradiation can be an efficient way of engineering the graphene edge with metal atoms. © 2012 The Royal Society of Chemistry.

Nano, Queensland and cryo-electron microscopy

International Nuclear Information System (INIS)

McDowall, A.W.

2002-01-01

Full text: In a recent review the authors, Wolfgang Baumeister and Alasdair Steven wrote, '....there is immense opportunity for Cryo-EM, especially as boosted by merging crystallographic structures of individual subunits into moderate resolution Cryo-EM density maps of whole complexes. Electron tomography has now advanced to the point where it is a realistic goal to glimpse molecular machines operating inside cells....' This statement gives testament to the advances made over the past 25 years by many labs around the world to the area of microscopy referred to as Cryo-EM and related 3-D computing technologies. Australian scientific societies have been eager followers of this progress and heard first hand of the new developments in the field at the 1984 ACEM-8 (2). Since those early days the ACEM and other Australian/NZ societies have sponsored numerous researchers and workshops in the field of Cryo-EM to their conferences, Helin Sabil, Wah Chiu, Ron Milligan, Richard Henderson and Werner Kuhlbrandt to name only a few. These visits have stimulated a desire from Australian/NZ researchers to establish collaborations and access to prominent labs in the USA and Europe, where the means and knowledge to provide Cryo EM and 3D reconstruction technology for studying macromolecular complexes is well established. However, Australia has not been backward in seeking to provide its home research community with access to a base in biological molecular microscopy and electron crystallography technology. Since the last ACEM we have seen the emergence of a number of crucial factors, which will make the establishment of a national research facility in this field an operational reality in early 2003. Well publicized is the development of Australia's newest and perhaps most unique research institute, the institute for Molecular Bioscience (IMB) to open at the University of Queensland (UQ) in 2002. The IMB will be the platform for a new research group in advanced computational 3D

Nanoscale Energy-Filtered Scanning Confocal Electron Microscopy Using a Double-Aberration-Corrected Transmission Electron Microscope

International Nuclear Information System (INIS)

Wang Peng; Behan, Gavin; Kirkland, Angus I.; Nellist, Peter D.; Takeguchi, Masaki; Hashimoto, Ayako; Mitsuishi, Kazutaka; Shimojo, Masayuki

2010-01-01

We demonstrate that a transmission electron microscope fitted with two spherical-aberration correctors can be operated as an energy-filtered scanning confocal electron microscope. A method for establishing this mode is described and initial results showing 3D chemical mapping with nanoscale sensitivity to height and thickness changes in a carbon film are presented. Importantly, uncorrected chromatic aberration does not limit the depth resolution of this technique and moreover performs an energy-filtering role, which is explained in terms of a combined depth and energy-loss response function.

Aberration-corrected imaging of active sites on industrial catalyst nanoparticles

DEFF Research Database (Denmark)

Gontard, Lionel Cervera; Chang, L-Y; Hetherington, CJD

2007-01-01

Picture perfect: Information about the local topologies of active sites on commercial nanoparticles can be gained with atomic resolution through spherical-aberration-corrected transmission electron microscopy (TEM). A powder of Pt nanoparticles on carbon black was examined with two advanced TEM t...

Aberration-corrected transmission electron microscopy analyses of GaAs/Si interfaces in wafer-bonded multi-junction solar cells

Energy Technology Data Exchange (ETDEWEB)

Häussler, Dietrich [Institute for Materials Science, Christian-Albrechts-University Kiel, Kaiserstraße 2, 24143 Kiel (Germany); Houben, Lothar [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Research Centre Juelich GmbH, 52425 Juelich (Germany); Essig, Stephanie [Fraunhofer Institute for Solar Energy Systems ISE, Heidenhofstraße 2, 79110 Freiburg (Germany); Kurttepeli, Mert [Institute for Materials Science, Christian-Albrechts-University Kiel, Kaiserstraße 2, 24143 Kiel (Germany); Dimroth, Frank [Fraunhofer Institute for Solar Energy Systems ISE, Heidenhofstraße 2, 79110 Freiburg (Germany); Dunin-Borkowski, Rafal E. [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Research Centre Juelich GmbH, 52425 Juelich (Germany); Jäger, Wolfgang, E-mail: wolfgang.jaeger@tf.uni-kiel.de [Institute for Materials Science, Christian-Albrechts-University Kiel, Kaiserstraße 2, 24143 Kiel (Germany)

2013-11-15

Aberration-corrected scanning transmission electron microscopy (STEM) and electron energy loss spectroscopy (EELS) investigations have been applied to investigate the structure and composition fluctuations near interfaces in wafer-bonded multi-junction solar cells. Multi-junction solar cells are of particular interest since efficiencies well above 40% have been obtained for concentrator solar cells which are based on III-V compound semiconductors. In this methodologically oriented investigation, we explore the potential of combining aberration-corrected high-angle annular dark-field STEM imaging (HAADF-STEM) with spectroscopic techniques, such as EELS and energy-dispersive X-ray spectroscopy (EDXS), and with high-resolution transmission electron microscopy (HR-TEM), in order to analyze the effects of fast atom beam (FAB) and ion beam bombardment (IB) activation treatments on the structure and composition of bonding interfaces of wafer-bonded solar cells on Si substrates. Investigations using STEM/EELS are able to measure quantitatively and with high precision the widths and the fluctuations in element distributions within amorphous interface layers of nanometer extensions, including those of light elements. Such measurements allow the control of the activation treatments and thus support assessing electrical conductivity phenomena connected with impurity and dopant distributions near interfaces for optimized performance of the solar cells. - Highlights: • Aberration-corrected TEM and EELS reveal structural and elemental profiles across GaAs/Si bond interfaces in wafer-bonded GaInP/GaAs/Si - multi-junction solar cells. • Fluctuations in elemental concentration in nanometer-thick amorphous interface layers, including the disrubutions of light elements, are measured using EELS. • The projected widths of the interface layers are determined on the atomic scale from STEM-HAADF measurements. • The effects of atom and ion beam activation treatment on the bonding

Present status and future prospects of spherical aberration corrected TEM/STEM for study of nanomaterials

International Nuclear Information System (INIS)

Tanaka, Nobuo

2008-01-01

The present status of Cs-corrected TEM/STEM is described from the viewpoint of the observation of nanomaterials. Characteristic features in TEM and STEM are explained using the experimental data obtained by our group and other research groups. Cs correction up to the 3rd-order aberration of an objective lens has already been established and research interest is focused on correcting the 5th-order spherical aberration and the chromatic aberration in combination with the development of a monochromator below an electron gun for smaller point-to-point resolution in optics. Another fundamental area of interest is the limitation of TEM and STEM resolution from the viewpoint of the scattering of electrons in crystals. The minimum size of the exit-wave function below samples undergoing TEM imaging is determined from the calculation of scattering around related atomic columns in the crystals. STEM does not have this limitation because the resolution is, in principle, determined by the probe size. One of the future prospects of Cs-corrected TEM/STEM is the possibility of extending the space around the sample holder by correcting the chromatic and spherical aberrations. This wider space will contribute to the ease of performing in situ experiments and various combinations of TEM and other analysis methods. High-resolution, in situ dynamic and 3D observations/analysis are the most important keywords in the next decade of high-resolution electron microscopy. (topical review)

Influence of Misalignment on High-Order Aberration Correction for Normal Human Eyes

Science.gov (United States)

Zhao, Hao-Xin; Xu, Bing; Xue, Li-Xia; Dai, Yun; Liu, Qian; Rao, Xue-Jun

2008-04-01

Although a compensation device can correct aberrations of human eyes, the effect will be degraded by its misalignment, especially for high-order aberration correction. We calculate the positioning tolerance of correction device for high-order aberrations, and within what degree the correcting effect is better than low-order aberration (defocus and astigmatism) correction. With fixed certain misalignment within the positioning tolerance, we calculate the residual wavefront rms aberration of the first-6 to first-35 terms along with the 3rd-5th terms of aberrations corrected, and the combined first-13 terms of aberrations are also studied under the same quantity of misalignment. However, the correction effect of high-order aberrations does not meliorate along with the increase of the high-order terms under some misalignment, moreover, some simple combined terms correction can achieve similar result as complex combinations. These results suggest that it is unnecessary to correct too much the terms of high-order aberrations which are difficult to accomplish in practice, and gives confidence to correct high-order aberrations out of the laboratory.

Influence of Misalignment on High-Order Aberration Correction for Normal Human Eyes

International Nuclear Information System (INIS)

Hao-Xin, Zhao; Bing, Xu; Li-Xia, Xue; Yun, Dai; Qian, Liu; Xue-Jun, Rao

2008-01-01

Although a compensation device can correct aberrations of human eyes, the effect will be degraded by its misalignment, especially for high-order aberration correction. We calculate the positioning tolerance of correction device for high-order aberrations, and within what degree the correcting effect is better than low-order aberration (defocus and astigmatism) correction. With fixed certain misalignment within the positioning tolerance, we calculate the residual wavefront rms aberration of the first-6 to first-35 terms along with the 3rd-5th terms of aberrations corrected, and the combined first-13 terms of aberrations are also studied under the same quantity of misalignment. However, the correction effect of high-order aberrations does not meliorate along with the increase of the high-order terms under some misalignment, moreover, some simple combined terms correction can achieve similar result as complex combinations. These results suggest that it is unnecessary to correct too much the terms of high-order aberrations which are difficult to accomplish in practice, and gives confidence to correct high-order aberrations out of the laboratory

First experimental proof for aberration correction in XPEEM: Resolution, transmission enhancement, and limitation by space charge effects

International Nuclear Information System (INIS)

Schmidt, Th.; Sala, A.; Marchetto, H.; Umbach, E.; Freund, H.-J.

2013-01-01

The positive effect of double aberration correction in x-ray induced Photoelectron Emission Microscopy (XPEEM) has been successfully demonstrated for both, the lateral resolution and the transmission, using the Au 4f XPS peak for element specific imaging at a kinetic energy of 113 eV. The lateral resolution is improved by a factor of four, compared to a non-corrected system, whereas the transmission is enhanced by a factor of 5 at a moderate resolution of 80 nm. With an optimized system setting, a lateral resolution of 18 nm could be achieved, which is up to now the best value reported for energy filtered XPEEM imaging. However, the absolute resolution does not yet reach the theoretical limit of 2 nm, which is due to space charge limitation. This occurs along the entire optical axis up to the contrast aperture. In XPEEM the pulsed time structure of the exciting soft x-ray light source causes a short and highly intense electron pulse, which results in an image blurring. In contrast, the imaging with elastically reflected electrons in the low energy electron microscopy (LEEM) mode yields a resolution clearly below 5 nm. Technical solutions to reduce the space charge effect in an aberration-corrected spectro-microscope are discussed. - Highlights: ► First successful double aberration correction in XPEEM. ► Improvement of resolution and transmission by aberration correction. ► Lateral resolution of 18 nm in energy filtered XPEEM is the best up to now reported value. ► First investigation of space charge effects in aberrations corrected PEEM

Aberration correction in photoemission microscopy and applications in photonics and plasmonics

Energy Technology Data Exchange (ETDEWEB)

Koenenkamp, Rolf [Portland State Univ., Portland, OR (United States)

2017-09-28

We report on the design, assembly, operation and application of an aberration-corrected photoemission electron microscope. The instrument used novel hyperbolic mirror-correctors with two and three electrodes that allowed simultaneous correction of spherical and chromatic aberrations. A spatial resolution of 5.4nm was obtained with this instrument in 2009, and 4.7nm in subsequent years. New imaging methodology was introduced involving interferometric imaging of light diffraction. This methodology was applied in nano-photonics and in the characterization of surface-plasmon polaritons. Photonic crystals and waveguides, optical antennas and new plasmonic devices such as routers, localizers and filters were designed and demonstrated using the new capabilities offered by the microscope.

Theoretical estimates of spherical and chromatic aberration in photoemission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Fitzgerald, J.P.S., E-mail: fit@pdx.edu; Word, R.C.; Könenkamp, R.

2016-01-15

We present theoretical estimates of the mean coefficients of spherical and chromatic aberration for low energy photoemission electron microscopy (PEEM). Using simple analytic models, we find that the aberration coefficients depend primarily on the difference between the photon energy and the photoemission threshold, as expected. However, the shape of the photoelectron spectral distribution impacts the coefficients by up to 30%. These estimates should allow more precise correction of aberration in PEEM in experimental situations where the aberration coefficients and precise electron energy distribution cannot be readily measured. - Highlights: • Spherical and chromatic aberration coefficients of the accelerating field in PEEM. • Compact, analytic expressions for coefficients depending on two emission parameters. • Effect of an aperture stop on the distribution is also considered.

Coordinate transformation based cryo-correlative methods for electron tomography and focused ion beam milling

International Nuclear Information System (INIS)

Fukuda, Yoshiyuki; Schrod, Nikolas; Schaffer, Miroslava; Feng, Li Rebekah; Baumeister, Wolfgang; Lucic, Vladan

2014-01-01

Correlative microscopy allows imaging of the same feature over multiple length scales, combining light microscopy with high resolution information provided by electron microscopy. We demonstrate two procedures for coordinate transformation based correlative microscopy of vitrified biological samples applicable to different imaging modes. The first procedure aims at navigating cryo-electron tomography to cellular regions identified by fluorescent labels. The second procedure, allowing navigation of focused ion beam milling to fluorescently labeled molecules, is based on the introduction of an intermediate scanning electron microscopy imaging step to overcome the large difference between cryo-light microscopy and focused ion beam imaging modes. These methods make it possible to image fluorescently labeled macromolecular complexes in their natural environments by cryo-electron tomography, while minimizing exposure to the electron beam during the search for features of interest. - Highlights: • Correlative light microscopy and focused ion beam milling of vitrified samples. • Coordinate transformation based cryo-correlative method. • Improved correlative light microscopy and cryo-electron tomography

Optimum aberration coefficients for recording high-resolution off-axis holograms in a Cs-corrected TEM

Energy Technology Data Exchange (ETDEWEB)

Linck, Martin, E-mail: linck@ceos-gmbh.de [CEOS GmbH, Englerstr. 28, D-69126 Heidelberg (Germany)

2013-01-15

Amongst the impressive improvements in high-resolution electron microscopy, the Cs-corrector also has significantly enhanced the capabilities of off-axis electron holography. Recently, it has been shown that the signal above noise in the reconstructable phase can be significantly improved by combining holography and hardware aberration correction. Additionally, with a spherical aberration close to zero, the traditional optimum focus for recording high-resolution holograms ('Lichte's defocus') has become less stringent and both, defocus and spherical aberration, can be selected freely within a certain range. This new degree of freedom can be used to improve the signal resolution in the holographically reconstructed object wave locally, e.g. at the atomic positions. A brute force simulation study for an aberration corrected 200 kV TEM is performed to determine optimum values for defocus and spherical aberration for best possible signal to noise in the reconstructed atomic phase signals. Compared to the optimum aberrations for conventional phase contrast imaging (NCSI), which produce 'bright atoms' in the image intensity, the resulting optimum values of defocus and spherical aberration for off-axis holography enable 'black atom contrast' in the hologram. However, they can significantly enhance the local signal resolution at the atomic positions. At the same time, the benefits of hardware aberration correction for high-resolution off-axis holography are preserved. It turns out that the optimum is depending on the object and its thickness and therefore not universal. -- Highlights: Black-Right-Pointing-Pointer Optimized aberration parameters for high-resolution off-axis holography. Black-Right-Pointing-Pointer Simulation and analysis of noise in high-resolution off-axis holograms. Black-Right-Pointing-Pointer Improving signal resolution in the holographically reconstructed phase shift. Black-Right-Pointing-Pointer Comparison of &apos

Optimum aberration coefficients for recording high-resolution off-axis holograms in a Cs-corrected TEM

International Nuclear Information System (INIS)

Linck, Martin

2013-01-01

Amongst the impressive improvements in high-resolution electron microscopy, the Cs-corrector also has significantly enhanced the capabilities of off-axis electron holography. Recently, it has been shown that the signal above noise in the reconstructable phase can be significantly improved by combining holography and hardware aberration correction. Additionally, with a spherical aberration close to zero, the traditional optimum focus for recording high-resolution holograms (“Lichte's defocus”) has become less stringent and both, defocus and spherical aberration, can be selected freely within a certain range. This new degree of freedom can be used to improve the signal resolution in the holographically reconstructed object wave locally, e.g. at the atomic positions. A brute force simulation study for an aberration corrected 200 kV TEM is performed to determine optimum values for defocus and spherical aberration for best possible signal to noise in the reconstructed atomic phase signals. Compared to the optimum aberrations for conventional phase contrast imaging (NCSI), which produce “bright atoms” in the image intensity, the resulting optimum values of defocus and spherical aberration for off-axis holography enable “black atom contrast” in the hologram. However, they can significantly enhance the local signal resolution at the atomic positions. At the same time, the benefits of hardware aberration correction for high-resolution off-axis holography are preserved. It turns out that the optimum is depending on the object and its thickness and therefore not universal. -- Highlights: ► Optimized aberration parameters for high-resolution off-axis holography. ► Simulation and analysis of noise in high-resolution off-axis holograms. ► Improving signal resolution in the holographically reconstructed phase shift. ► Comparison of “black” and “white” atom contrast in off-axis holograms.

TOPICAL REVIEW: Present status and future prospects of spherical aberration corrected TEM/STEM for study of nanomaterials

Directory of Open Access Journals (Sweden)

Nobuo Tanaka

2008-01-01

Full Text Available The present status of Cs-corrected TEM/STEM is described from the viewpoint of the observation of nanomaterials. Characteristic features in TEM and STEM are explained using the experimental data obtained by our group and other research groups. Cs correction up to the 3rd-order aberration of an objective lens has already been established and research interest is focused on correcting the 5th-order spherical aberration and the chromatic aberration in combination with the development of a monochromator below an electron gun for smaller point-to-point resolution in optics. Another fundamental area of interest is the limitation of TEM and STEM resolution from the viewpoint of the scattering of electrons in crystals. The minimum size of the exit-wave function below samples undergoing TEM imaging is determined from the calculation of scattering around related atomic columns in the crystals. STEM does not have this limitation because the resolution is, in principle, determined by the probe size. One of the future prospects of Cs-corrected TEM/STEM is the possibility of extending the space around the sample holder by correcting the chromatic and spherical aberrations. This wider space will contribute to the ease of performing in situ experiments and various combinations of TEM and other analysis methods. High-resolution, in situ dynamic and 3D observations/analysis are the most important keywords in the next decade of high-resolution electron microscopy.

Parainfluenza virus type 5 (PIV-5) morphology revealed by cryo-electron microscopy.

Science.gov (United States)

Terrier, Olivier; Rolland, Jean-Paul; Rosa-Calatrava, Manuel; Lina, Bruno; Thomas, Daniel; Moules, Vincent

2009-06-01

The knowledge of parainfluenza type 5 (PIV-5) virion morphology is essentially based on the observation of negatively stained preparations in conventional transmission electron microscopy (CTEM). In this study, the ultrastructure of frozen-hydrated intact PIV-5 was examined by cryo-electron microscopy (cryo-EM). Cryo-EM revealed a majority of spherical virions (70%), with a lower pleiomorphy than originally observed in CTEM. Phospholipid bilayer thickness, spike length and glycoprotein spikes density were measured. About 2000 glycoprotein spikes were present in an average-sized spherical virion. Altogether, these data depict a more precise view of PIV-5 morphology.

Prospects for versatile phase manipulation in the TEM: Beyond aberration correction

International Nuclear Information System (INIS)

Guzzinati, Giulio; Clark, Laura; Béché, Armand; Juchtmans, Roeland; Van Boxem, Ruben; Mazilu, Michael; Verbeeck, Jo

2015-01-01

In this paper we explore the desirability of a transmission electron microscope in which the phase of the electron wave can be freely controlled. We discuss different existing methods to manipulate the phase of the electron wave and their limitations. We show how with the help of current techniques the electron wave can already be crafted into specific classes of waves each having their own peculiar properties. Assuming a versatile phase modulation device is feasible, we explore possible benefits and methods that could come into existence borrowing from light optics where the so-called spatial light modulators provide programmable phase plates for quite some time now. We demonstrate that a fully controllable phase plate building on Harald Rose's legacy in aberration correction and electron optics in general would open an exciting field of research and applications. - Highlights: • We offer a review of available phase manipulation techniques. • We demonstrate a method for producing Airy waves through aberration manipulation. • We outline hypothetical applications of arbitrary phase manipulation methods

Zernike phase contrast cryo-electron tomography of whole bacterial cells.

Science.gov (United States)

Guerrero-Ferreira, Ricardo C; Wright, Elizabeth R

2014-01-01

Cryo-electron tomography (cryo-ET) provides three-dimensional (3D) structural information of bacteria preserved in a native, frozen-hydrated state. The typical low contrast of tilt-series images, a result of both the need for a low electron dose and the use of conventional defocus phase-contrast imaging, is a challenge for high-quality tomograms. We show that Zernike phase-contrast imaging allows the electron dose to be reduced. This limits movement of gold fiducials during the tilt series, which leads to better alignment and a higher-resolution reconstruction. Contrast is also enhanced, improving visibility of weak features. The reduced electron dose also means that more images at more tilt angles could be recorded, further increasing resolution. Copyright © 2013 Elsevier Inc. All rights reserved.

Pulse compressor with aberration correction

Energy Technology Data Exchange (ETDEWEB)

Mankos, Marian [Electron Optica, Inc., Palo Alto, CA (United States)

2015-11-30

In this SBIR project, Electron Optica, Inc. (EOI) is developing an electron mirror-based pulse compressor attachment to new and retrofitted dynamic transmission electron microscopes (DTEMs) and ultrafast electron diffraction (UED) cameras for improving the temporal resolution of these instruments from the characteristic range of a few picoseconds to a few nanoseconds and beyond, into the sub-100 femtosecond range. The improvement will enable electron microscopes and diffraction cameras to better resolve the dynamics of reactions in the areas of solid state physics, chemistry, and biology. EOI’s pulse compressor technology utilizes the combination of electron mirror optics and a magnetic beam separator to compress the electron pulse. The design exploits the symmetry inherent in reversing the electron trajectory in the mirror in order to compress the temporally broadened beam. This system also simultaneously corrects the chromatic and spherical aberration of the objective lens for improved spatial resolution. This correction will be found valuable as the source size is reduced with laser-triggered point source emitters. With such emitters, it might be possible to significantly reduce the illuminated area and carry out ultrafast diffraction experiments from small regions of the sample, e.g. from individual grains or nanoparticles. During phase I, EOI drafted a set of candidate pulse compressor architectures and evaluated the trade-offs between temporal resolution and electron bunch size to achieve the optimum design for two particular applications with market potential: increasing the temporal and spatial resolution of UEDs, and increasing the temporal and spatial resolution of DTEMs. Specialized software packages that have been developed by MEBS, Ltd. were used to calculate the electron optical properties of the key pulse compressor components: namely, the magnetic prism, the electron mirror, and the electron lenses. In the final step, these results were folded

Aberration corrected and 3D cryo-tomography HAADF-STEM surface studies of ZnO tetrapods

International Nuclear Information System (INIS)

Ward, M R; Gai, P L; Boyes, E D; Sugiura, H; Tanaka, N; Yoshida, K

2012-01-01

We present a morphology study of ZnO tetrapods using aberration corrected TEM, HAADF-STEM and 3D HAADF-STEM cryotomography as an alternative to more conventional TEM and SEM techniques. We use 3D IMOD reconstructions to show that the {11-bar 0} facets dominate the total surface area of uniform hexagonal prism tetrapods. Using HRTEM we show that the small tetrapods have a zincblende phase core from which the four legs extend. The facets and the edges of these legs were found to be atomically clean and flat with the potential for ZnO tetrapods as model substrates. We deposited ultrafine Pt/Pd nanoparticles onto the tetrapods and investigated the resulting morphologies. We found using HAADF-STEM cryotomography and reconstruction techniques that the nanoparticle coverage gave separate nanoparticles and overall uniform coverage. We believe these techniques and the results from them could be useful for the development of nanoparticle-ZnO tetrapod composite systems with applications in optoelectronics, gas sensing and catalysis.

The CryoSat Interferometer after 6 years in orbit: calibration and achievable performance

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; De Bartolomei, Maurizio; Bouffard, Jerome; Parrinello, Tommaso

2016-04-01

The main payload of CryoSat is a Ku-band pulse width limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter). When commanded in SARIn (synthetic aperture radar interferometry) mode, through coherent along-track processing of the returns received from two antennas, the interferometric phase related to the first arrival of the echo is used to retrieve the angle of arrival of the scattering in the across-track direction. In fact, the across-track echo direction can be derived by exploiting the precise knowledge of the baseline vector (i.e. the vector between the two antennas centers of phase) and simple geometry. The end-to-end calibration strategy for the CryoSat interferometer consists on in-orbit calibration campaigns following the approach described in [1]. From the beginning of the CryoSat mission, about once a year the interferometer calibration campaigns have been periodically performed by rolling left and right the spacecraft of about ±0.4 deg. This abstract is aimed at presenting our analysis of the calibration parameters and of the achievable performance of the CryoSat interferometer over the 6 years of mission. Additionally, some further studies have been performed to assess the accuracy of the roll angle computed on ground as function of the aberration (the apparent displacement of a celestial object from its true position, caused by the relative motion of the observer and the object) correction applied to the attitude quaternions, provided by the Star Tracker mounted on-board. In fact, being the roll information crucial to obtain an accurate estimate of the angle of arrival, the data from interferometer calibration campaigns have been used to verify how the application of the aberration correction affects the roll information and, in turns, the measured angle of arrival. [1] Galin, N.; Wingham, D.J.; Cullen, R.; Fornari, M.; Smith, W.H.F.; Abdalla, S., "Calibration of the CryoSat-2 Interferometer and Measurement of Across

Dynamic Correction of Higher-Order Deflection Aberrations in the Environmental SEM

Czech Academy of Sciences Publication Activity Database

Oral, Martin; Neděla, Vilém

2015-01-01

Roč. 21, S4 (2015), s. 194-199 ISSN 1431-9276 R&D Projects: GA ČR(CZ) GA14-22777S; GA MŠk(CZ) LO1212 Institutional support: RVO:68081731 Keywords : environmental SEM * ESEM * shifted deflection pivot point * Higher order deflection aberrations * vignetting * dynamic focusing * dynamic stigmator * dynamic correction Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.730, year: 2015

Chromatic Aberration Correction for Atomic Resolution TEM Imaging from 20 to 80 kV.

Science.gov (United States)

Linck, Martin; Hartel, Peter; Uhlemann, Stephan; Kahl, Frank; Müller, Heiko; Zach, Joachim; Haider, Max; Niestadt, Marcel; Bischoff, Maarten; Biskupek, Johannes; Lee, Zhongbo; Lehnert, Tibor; Börrnert, Felix; Rose, Harald; Kaiser, Ute

2016-08-12

Atomic resolution in transmission electron microscopy of thin and light-atom materials requires a rigorous reduction of the beam energy to reduce knockon damage. However, at the same time, the chromatic aberration deteriorates the resolution of the TEM image dramatically. Within the framework of the SALVE project, we introduce a newly developed C_{c}/C_{s} corrector that is capable of correcting both the chromatic and the spherical aberration in the range of accelerating voltages from 20 to 80 kV. The corrector allows correcting axial aberrations up to fifth order as well as the dominating off-axial aberrations. Over the entire voltage range, optimum phase-contrast imaging conditions for weak signals from light atoms can be adjusted for an optical aperture of at least 55 mrad. The information transfer within this aperture is no longer limited by chromatic aberrations. We demonstrate the performance of the microscope using the examples of 30 kV phase-contrast TEM images of graphene and molybdenum disulfide, showing unprecedented contrast and resolution that matches image calculations.

Compensation of aberrations of deflected electron probe by means of dynamical focusing with stigmator

International Nuclear Information System (INIS)

Baba, Norio; Ebe, Toyoe; Ikehata, Koichi; Ito, Yasuhiro; Terada, Hiroshi

1979-01-01

Electron beam passing through a deflecting field is in general, subjected to aberrations such as distortion, astigmatism and coma in accordance with the deflecting angle. Accordingly the aberration defect of deflected beam is the most serious limiting factor in the performances of micromachining, microminiaturization and high resolution scanning electron microscopes. From many investigators' results, it is obvious that three important compensation methods to aberrations exist in principle, i.e., double deflection system, dynamical focusing, and the dynamical correction using a stigmator. In this paper, based on the aberration formula derived from the eikonal or the path method, the practical data of the aberration constants of deflected electron beam for the sequential deflection system with parallel plates are calculated, and using its result, the distorted spot patterns of an electron probe deflected in two-dimensional directions for various defocusings are graphically displayed by the aid of a computer. Further, by means of the dynamical focusing with a stigmator, the conditions to completely compensate the second order astigmatic aberration are derived, and spot patterns and the electron density distributions within the spots in the case when the compensating conditions are satisfied are also graphically displayed. (Wakatsuki, Y.)

National Cryo-Electron Microscopy Facility

Science.gov (United States)

Information about the National Cryo-EM Facility at NCI, created to provide researchers access to the latest cryo-EM technology for high resolution imaging. Includes timeline for installation and how to access the facility.

Single-particle cryo-electron microscopy of Rift Valley fever virus

OpenAIRE

Sherman, Michael B.; Freiberg, Alexander N.; Holbrook, Michael R.; Watowich, Stanley J.

2009-01-01

Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human veterinary pathogen causing acute hepatitis in ruminants and has the potential to Single-particle cryo-EM reconstruction of RVFV MP-12 hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on...

Sextupole system for the correction of spherical aberration

Science.gov (United States)

Crewe, A.V.; Kopf, D.A.

In an electron beam device in which an electron beam is developed and then focused by a lens to a particular spot, there is provided a means for eliminating spherical aberration. A sextupole electromagnetic lens is positioned between two focusing lenses. The interaction of the sextupole with the beam compensates for spherical aberration. (GHT)

Spatial and temporal resolution in cryo-electron microscopy : a scope for nano-chemistry

NARCIS (Netherlands)

Frederik, P.M.; Sommerdijk, N.A.J.M.

2005-01-01

Cryo-electron microscopy has evolved in an established approach to study the structure of bio-colloids. Recent developments in instrumentation and automation, often demanded by life sciences, made cryo-EM a general tool in colloid chemistry. Recently improved instrumentation for vitrification has

Correcting the Chromatic Aberration in Barrel Distortion of Endoscopic Images

Directory of Open Access Journals (Sweden)

Y. M. Harry Ng

2003-04-01

Full Text Available Modern endoscopes offer physicians a wide-angle field of view (FOV for minimally invasive therapies. However, the high level of barrel distortion may prevent accurate perception of image. Fortunately, this kind of distortion may be corrected by digital image processing. In this paper we investigate the chromatic aberrations in the barrel distortion of endoscopic images. In the past, chromatic aberration in endoscopes is corrected by achromatic lenses or active lens control. In contrast, we take a computational approach by modifying the concept of image warping and the existing barrel distortion correction algorithm to tackle the chromatic aberration problem. In addition, an error function for the determination of the level of centroid coincidence is proposed. Simulation and experimental results confirm the effectiveness of our method.

Optical depth sectioning in the aberration-corrected scanning transmission and scanning confocal electron microscope

International Nuclear Information System (INIS)

Behan, G; Nellist, P D

2008-01-01

The use of spherical aberration correctors in the scanning transmission electron microscope (STEM) has the effect of reducing the depth of field of the microscope, making three-dimensional imaging of a specimen possible by optical sectioning. Depth resolution can be improved further by placing aberration correctors and lenses pre and post specimen to achieve an imaging mode known as scanning confocal electron microscopy (SCEM). We present the calculated incoherent point spread functions (PSF) and optical transfer functions (OTF) of a STEM and SCEM. The OTF for a STEM is shown to have a missing cone region which results in severe blurring along the optic axis, which can be especially severe for extended objects. We also present strategies for reconstruction of experimental data, such as three-dimensional deconvolution of the point spread function.

Towards atomic scale engineering of rare-earth-doped SiAlON ceramics through aberration-corrected scanning transmission electron microscopy

International Nuclear Information System (INIS)

Yurdakul, Hilmi; Idrobo, Juan C.; Pennycook, Stephen J.; Turan, Servet

2011-01-01

Direct visualization of rare earths in α- and β-SiAlON unit-cells is performed through Z-contrast imaging technique in an aberration-corrected scanning transmission electron microscope. The preferential occupation of Yb and Ce atoms in different interstitial locations of β-SiAlON lattice is demonstrated, yielding higher solubility for Yb than Ce. The triangular-like host sites in α-SiAlON unit cell accommodate more Ce atoms than hexagonal sites in β-SiAlON. We think that our results will be applicable as guidelines for many kinds of rare-earth-doped materials.

Identification of light elements in silicon nitride by aberration-corrected scanning transmission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Idrobo, Juan C., E-mail: idrobojc@ornl.gov [Materials Science and Technology Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831 (United States); Walkosz, Weronika [Materials Science Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Department of Physics, University of Illinois at Chicago, Chicago, IL 60607 (United States); Klie, Robert F.; Oeguet, Serdar [Department of Physics, University of Illinois at Chicago, Chicago, IL 60607 (United States)

2012-12-15

In silicon nitride structural ceramics, the overall mechanical and thermal properties are controlled by the atomic and electronic structures at the interface between the ceramic grains and the amorphous intergranular films (IGFs) formed by various sintering additives. In the last ten years the atomic arrangements of heavy elements (rare-earths) at the Si{sub 3}N{sub 4}/IGF interfaces have been resolved. However, the atomic position of light elements, without which it is not possible to obtain a complete description of the interfaces, has been lacking. This review article details the authors' efforts to identify the atomic arrangement of light elements such as nitrogen and oxygen at the Si{sub 3}N{sub 4}/SiO{sub 2} interface and in bulk Si{sub 3}N{sub 4} using aberration-corrected scanning transmission electron microscopy. -- Highlights: Black-Right-Pointing-Pointer Revealing the atomic structure of the {alpha}-Si{sub 3}N{sub 4}/SiO{sub 2} interface. Black-Right-Pointing-Pointer Identification and lattice location of oxygen impurities in bulk {alpha}-Si{sub 3}N{sub 4}. Black-Right-Pointing-Pointer Short range ordering of nitrogen and oxygen at the {beta}-Si{sub 3}N{sub 4}/SiO{sub 2} interface.

A cylindrical specimen holder for electron cryo-tomography

International Nuclear Information System (INIS)

Palmer, Colin M.; Löwe, Jan

2014-01-01

The use of slab-like flat specimens for electron cryo-tomography restricts the range of viewing angles that can be used. This leads to the “missing wedge” problem, which causes artefacts and anisotropic resolution in reconstructed tomograms. Cylindrical specimens provide a way to eliminate the problem, since they allow imaging from a full range of viewing angles around the tilt axis. Such specimens have been used before for tomography of radiation-insensitive samples at room temperature, but never for frozen-hydrated specimens. Here, we demonstrate the use of thin-walled carbon tubes as specimen holders, allowing the preparation of cylindrical frozen-hydrated samples of ribosomes, liposomes and whole bacterial cells. Images acquired from these cylinders have equal quality at all viewing angles, and the accessible tilt range is restricted only by the physical limits of the microscope. Tomographic reconstructions of these specimens demonstrate that the effects of the missing wedge are substantially reduced, and could be completely eliminated if a full tilt range was used. The overall quality of these tomograms is still lower than that obtained by existing methods, but improvements are likely in future. - Highlights: • The missing wedge is a serious problem for electron cryo-tomography. • Cylindrical specimens allow the missing wedge to be eliminated. • Carbon nanopipettes can be used as cylindrical holders for tomography of frozen-hydrated specimens. • Cryo-tomography of cylindrical biological samples demonstrates a reduction of deleterious effects associated with the missing wedge

Aberration corrected and monochromated environmental transmission electron microscopy: challenges and prospects for materials science

DEFF Research Database (Denmark)

Hansen, Thomas Willum; Wagner, Jakob Birkedal; Dunin-Borkowski, Rafal E.

2010-01-01

The latest generation of environmental transmission electron microscopes incorporates aberration correctors and monochromators, allowing studies of chemical reactions and growth processes with improved spatial resolution and spectral sensitivity. Here, we describe the performance of such an instr...

A simple cryo-holder facilitates specimen observation under a conventional scanning electron microscope.

Science.gov (United States)

Tang, Chih-Yuan; Huang, Rong-Nan; Kuo-Huang, Ling-Long; Kuo, Tai-Chih; Yang, Ya-Yun; Lin, Ching-Yeh; Jane, Wann-Neng; Chen, Shiang-Jiuun

2012-02-01

A pre-cryogenic holder (cryo-holder) facilitating cryo-specimen observation under a conventional scanning electron microscope (SEM) is described. This cryo-holder includes a specimen-holding unit (the stub) and a cryogenic energy-storing unit (a composite of three cylinders assembled with a screw). After cooling, the cryo-holder can continue supplying cryogenic energy to extend the observation time for the specimen in a conventional SEM. Moreover, the cryogenic energy-storing unit could retain appropriate liquid nitrogen that can evaporate to prevent frost deposition on the surface of the specimen. This device is proved feasible for various tissues and cells, and can be applied to the fields of both biology and material science. We have employed this novel cryo-holder for observation of yeast cells, trichome, and epidermal cells in the leaf of Arabidopsis thaliana, compound eyes of insects, red blood cells, filiform papillae on the surface of rat tongue, agar medium, water molecules, penicillium, etc. All results suggested that the newly designed cryo-holder is applicable for cryo-specimen observation under a conventional SEM without cooling system. Most importantly, the design of this cryo-holder is simple and easy to operate and could adapt a conventional SEM to a plain type cryo-SEM affordable for most laboratories. Copyright © 2011 Wiley Periodicals, Inc.

Rapid increase of near atomic resolution virus capsid structures determined by cryo-electron microscopy.

Science.gov (United States)

Ho, Phuong T; Reddy, Vijay S

2018-01-01

The recent technological advances in electron microscopes, detectors, as well as image processing and reconstruction software have brought single particle cryo-electron microscopy (cryo-EM) into prominence for determining structures of bio-molecules at near atomic resolution. This has been particularly true for virus capsids, ribosomes, and other large assemblies, which have been the ideal specimens for structural studies by cryo-EM approaches. An analysis of time series metadata of virus structures on the methods of structure determination, resolution of the structures, and size of the virus particles revealed a rapid increase in the virus structures determined by cryo-EM at near atomic resolution since 2010. In addition, the data highlight the median resolution (∼3.0 Å) and size (∼310.0 Å in diameter) of the virus particles determined by X-ray crystallography while no such limits exist for cryo-EM structures, which have a median diameter of 508 Å. Notably, cryo-EM virus structures in the last four years have a median resolution of 3.9 Å. Taken together with minimal sample requirements, not needing diffraction quality crystals, and being able to achieve similar resolutions of the crystal structures makes cryo-EM the method of choice for current and future virus capsid structure determinations. Copyright © 2017 Elsevier Inc. All rights reserved.

Effect of correction of aberration dynamics on chaos in human ocular accommodation.

Science.gov (United States)

Hampson, Karen M; Cufflin, Matthew P; Mallen, Edward A H

2013-11-15

We used adaptive optics to determine the effect of monochromatic aberration dynamics on the level of chaos in the accommodation control system. Four participants viewed a stationary target while the dynamics of their aberrations were either left uncorrected, defocus was corrected, or all aberrations except defocus were corrected. Chaos theory analysis was used to discern changes in the accommodative microfluctuations. We found a statistically significant reduction in the chaotic nature of the accommodation microfluctuations during correction of defocus, but not when all aberrations except defocus were corrected. The Lyapunov exponent decreased from 0.71 ± 0.07 D/s (baseline) to 0.55 ± 0.03 D/s (correction of defocus fluctuations). As the reduction of chaos in physiological signals is indicative of stress to the system, the results indicate that for the participants included in this study, fluctuations in defocus have a more profound effect than those of the other aberrations. There were no changes in the power spectrum between experimental conditions. Hence chaos theory analysis is a more subtle marker of changes in the accommodation control system and will be of value in the study of myopia onset and progression.

Numerical correction of anti-symmetric aberrations in single HRTEM images of weakly scattering 2D-objects

International Nuclear Information System (INIS)

Lehtinen, Ossi; Geiger, Dorin; Lee, Zhongbo; Whitwick, Michael Brian; Chen, Ming-Wei; Kis, Andras; Kaiser, Ute

2015-01-01

Here, we present a numerical post-processing method for removing the effect of anti-symmetric residual aberrations in high-resolution transmission electron microscopy (HRTEM) images of weakly scattering 2D-objects. The method is based on applying the same aberrations with the opposite phase to the Fourier transform of the recorded image intensity and subsequently inverting the Fourier transform. We present the theoretical justification of the method, and its verification based on simulated images in the case of low-order anti-symmetric aberrations. Ultimately the method is applied to experimental hardware aberration-corrected HRTEM images of single-layer graphene and MoSe 2 resulting in images with strongly reduced residual low-order aberrations, and consequently improved interpretability. Alternatively, this method can be used to estimate by trial and error the residual anti-symmetric aberrations in HRTEM images of weakly scattering objects

Spherical aberration correction with threefold symmetric line currents.

Science.gov (United States)

Hoque, Shahedul; Ito, Hiroyuki; Nishi, Ryuji; Takaoka, Akio; Munro, Eric

2016-02-01

It has been shown that N-fold symmetric line current (henceforth denoted as N-SYLC) produces 2N-pole magnetic fields. In this paper, a threefold symmetric line current (N3-SYLC in short) is proposed for correcting 3rd order spherical aberration of round lenses. N3-SYLC can be realized without using magnetic materials, which makes it free of the problems of hysteresis, inhomogeneity and saturation. We investigate theoretically the basic properties of an N3-SYLC configuration which can in principle be realized by simple wires. By optimizing the parameters of a system with beam energy of 5.5keV, the required excitation current for correcting 3rd order spherical aberration coefficient of 400 mm is less than 1AT, and the residual higher order aberrations can be kept sufficiently small to obtain beam size of less than 1 nm for initial slopes up to 5 mrad. Copyright © 2015 Elsevier B.V. All rights reserved.

Correcting the wavefront aberration of membrane mirror based on liquid crystal spatial light modulator

Science.gov (United States)

Yang, Bin; Wei, Yin; Chen, Xinhua; Tang, Minxue

2014-11-01

Membrane mirror with flexible polymer film substrate is a new-concept ultra lightweight mirror for space applications. Compared with traditional mirrors, membrane mirror has the advantages of lightweight, folding and deployable, low cost and etc. Due to the surface shape of flexible membrane mirror is easy to deviate from the design surface shape, it will bring wavefront aberration to the optical system. In order to solve this problem, a method of membrane mirror wavefront aberration correction based on the liquid crystal spatial light modulator (LCSLM) will be studied in this paper. The wavefront aberration correction principle of LCSLM is described and the phase modulation property of a LCSLM is measured and analyzed firstly. Then the membrane mirror wavefront aberration correction system is designed and established according to the optical properties of a membrane mirror. The LCSLM and a Hartmann-Shack sensor are used as a wavefront corrector and a wavefront detector, respectively. The detected wavefront aberration is calculated and converted into voltage value on LCSLM for the mirror wavefront aberration correction by programming in Matlab. When in experiment, the wavefront aberration of a glass plane mirror with a diameter of 70 mm is measured and corrected for verifying the feasibility of the experiment system and the correctness of the program. The PV value and RMS value of distorted wavefront are reduced and near diffraction limited optical performance is achieved. On this basis, the wavefront aberration of the aperture center Φ25 mm in a membrane mirror with a diameter of 200 mm is corrected and the errors are analyzed. It provides a means of correcting the wavefront aberration of membrane mirror.

Chromatic aberration correction: an enhancement to the calibration of low-cost digital dermoscopes.

Science.gov (United States)

Wighton, Paul; Lee, Tim K; Lui, Harvey; McLean, David; Atkins, M Stella

2011-08-01

We present a method for calibrating low-cost digital dermoscopes that corrects for color and inconsistent lighting and also corrects for chromatic aberration. Chromatic aberration is a form of radial distortion that often occurs in inexpensive digital dermoscopes and creates red and blue halo-like effects on edges. Being radial in nature, distortions due to chromatic aberration are not constant across the image, but rather vary in both magnitude and direction. As a result, distortions are not only visually distracting but could also mislead automated characterization techniques. Two low-cost dermoscopes, based on different consumer-grade cameras, were tested. Color is corrected by imaging a reference and applying singular value decomposition to determine the transformation required to ensure accurate color reproduction. Lighting is corrected by imaging a uniform surface and creating lighting correction maps. Chromatic aberration is corrected using a second-order radial distortion model. Our results for color and lighting calibration are consistent with previously published results, while distortions due to chromatic aberration can be reduced by 42-47% in the two systems considered. The disadvantages of inexpensive dermoscopy can be quickly substantially mitigated with a suitable calibration procedure. © 2011 John Wiley & Sons A/S.

Low-energy foil aberration corrector

International Nuclear Information System (INIS)

Aken, R.H. van; Hagen, C.W.; Barth, J.E.; Kruit, P.

2002-01-01

A spherical and chromatic aberration corrector for electron microscopes is proposed, consisting of a thin foil sandwiched between two apertures. The electrons are retarded at the foil to almost zero energy, so that they can travel ballistically through the foil. It is shown that such a low-voltage corrector has a negative spherical aberration for not too large distances between aperture and foil, as well as a negative chromatic aberration. For various distances the third- and fifth-order spherical aberration coefficients and the first- and second-order chromatic aberration coefficients are calculated using ray tracing. Provided that the foils have sufficient electron transmission the corrector is able to correct the third-order spherical aberration and the first-order chromatic aberration of a typical low-voltage scanning electron microscope. Preliminary results show that the fifth-order spherical aberration and the second-order chromatic aberration can be kept sufficiently low

In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate

Energy Technology Data Exchange (ETDEWEB)

Barton, B.; Rhinow, D.; Walter, A.; Schroeder, R. [Max Planck Institute of Biophysics, Max-von-Laue Str. 3, 60438 Frankfurt am Main (Germany); Benner, G.; Majorovits, E.; Matijevic, M.; Niebel, H. [Carl Zeiss NTS GmbH, D-73447 Oberkochen (Germany); Mueller, H.; Haider, M. [CEOS GmbH, Englerstr. 26, 69126 Heidleberg (Germany); Lacher, M.; Schmitz, S.; Holik, P. [Caesar Research Center, Ludwig-Erhard-Allee 2, D-53175 Bonn (Germany); Kuehlbrandt, W., E-mail: werner.kuehlbrandt@mpibp-frankfurt.mpg.de [Max Planck Institute of Biophysics, Max-von-Laue Str. 3, 60438 Frankfurt am Main (Germany)

2011-12-15

We report the implementation of an electrostatic Einzel lens (Boersch) phase plate in a prototype transmission electron microscope dedicated to aberration-corrected cryo-EM. The combination of phase plate, C{sub s} corrector and Diffraction Magnification Unit (DMU) as a new electron-optical element ensures minimal information loss due to obstruction by the phase plate and enables in-focus phase contrast imaging of large macromolecular assemblies. As no defocussing is necessary and the spherical aberration is corrected, maximal, non-oscillating phase contrast transfer can be achieved up to the information limit of the instrument. A microchip produced by a scalable micro-fabrication process has 10 phase plates, which are positioned in a conjugate, magnified diffraction plane generated by the DMU. Phase plates remained fully functional for weeks or months. The large distance between phase plate and the cryo sample permits the use of an effective anti-contaminator, resulting in ice contamination rates of <0.6 nm/h at the specimen. Maximal in-focus phase contrast was obtained by applying voltages between 80 and 700 mV to the phase plate electrode. The phase plate allows for in-focus imaging of biological objects with a signal-to-noise of 5-10 at a resolution of 2-3 nm, as demonstrated for frozen-hydrated virus particles and purple membrane at liquid-nitrogen temperature. -- Highlights: Black-Right-Pointing-Pointer We implement an electrostatic Boersch phase plate into a dedicated prototypical TEM. Black-Right-Pointing-Pointer Phase contrast aberration-corrected electron microscope (PACEM) includes a diffraction magnification unit (DMU). Black-Right-Pointing-Pointer DMU minimizes obstruction of low spatial frequencies by the phase plate. Black-Right-Pointing-Pointer In-focus phase contrast generation is demonstrated for frozen-hydrated biological specimens.

3D structure of eukaryotic flagella/cilia by cryo-electron tomography.

Science.gov (United States)

Ishikawa, Takashi

2013-01-01

Flagella/cilia are motile organelles with more than 400 proteins. To understand the mechanism of such complex systems, we need methods to describe molecular arrange-ments and conformations three-dimensionally in vivo. Cryo-electron tomography enabled us such a 3D structural analysis. Our group has been working on 3D structure of flagella/cilia using this method and revealed highly ordered and beautifully organized molecular arrangement. 3D structure gave us insights into the mechanism to gener-ate bending motion with well defined waveforms. In this review, I summarize our recent structural studies on fla-gella/cilia by cryo-electron tomography, mainly focusing on dynein microtubule-based ATPase motor proteins and the radial spoke, a regulatory protein complex.

Imaging endosomes and autophagosomes in whole mammalian cells using correlative cryo-fluorescence and cryo-soft X-ray microscopy (cryo-CLXM)

International Nuclear Information System (INIS)

Duke, Elizabeth M.H.; Razi, Minoo; Weston, Anne; Guttmann, Peter; Werner, Stephan; Henzler, Katja; Schneider, Gerd; Tooze, Sharon A.; Collinson, Lucy M.

2014-01-01

Cryo-soft X-ray tomography (cryo-SXT) is a powerful imaging technique that can extract ultrastructural information from whole, unstained mammalian cells as close to the living state as possible. Subcellular organelles including the nucleus, the Golgi apparatus and mitochondria have been identified by morphology alone, due to the similarity in contrast to transmission electron micrographs. In this study, we used cryo-SXT to image endosomes and autophagosomes, organelles that are particularly susceptible to chemical fixation artefacts during sample preparation for electron microscopy. We used two approaches to identify these compartments. For early and recycling endosomes, which are accessible to externally-loaded markers, we used an anti-transferrin receptor antibody conjugated to 10 nm gold particles. For autophagosomes, which are not accessible to externally-applied markers, we developed a correlative cryo-fluorescence and cryo-SXT workflow (cryo-CLXM) to localise GFP-LC3 and RFP-Atg9. We used a stand-alone cryo-fluorescence stage in the home laboratory to localise the cloned fluorophores, followed by cryo-soft X-ray tomography at the synchrotron to analyse cellular ultrastructure. We mapped the 3D ultrastructure of the endocytic and autophagic structures, and discovered clusters of omegasomes arising from ‘hotspots’ on the ER. Thus, immunogold markers and cryo-CLXM can be used to analyse cellular processes that are inaccessible using other imaging modalities. - Highlights: • We image whole, unstained mammalian cells using cryo-soft X-ray tomography. • Endosomes are identified using a gold marker for the transferrin receptor. • A new workflow for correlative cryo-fluorescence and cryo-SXT is used to locate early autophagosomes. • Interactions between endosomes, endoplasmic reticulum and forming autophagosomes are mapped in 3D. • Multiple omegasomes are shown to form at ‘hotspots’ on the endoplasmic reticulum

Three-dimensional structural analysis of eukaryotic flagella/cilia by electron cryo-tomography

International Nuclear Information System (INIS)

Bui, Khanh Huy; Pigino, Gaia; Ishikawa, Takashi

2011-01-01

Based on the molecular architecture revealed by electron cryo-tomography, the mechanism of the bending motion of eukaryotic flagella/cilia is discussed. Electron cryo-tomography is a potential approach to analyzing the three-dimensional conformation of frozen hydrated biological macromolecules using electron microscopy. Since projections of each individual object illuminated from different orientations are merged, electron tomography is capable of structural analysis of such heterogeneous environments as in vivo or with polymorphism, although radiation damage and the missing wedge are severe problems. Here, recent results on the structure of eukaryotic flagella, which is an ATP-driven bending organelle, from green algae Chlamydomonas are presented. Tomographic analysis reveals asymmetric molecular arrangements, especially that of the dynein motor proteins, in flagella, giving insight into the mechanism of planar asymmetric bending motion. Methodological challenges to obtaining higher-resolution structures from this technique are also discussed

A lateral chromatic aberration correction system for ultrahigh-definition color video camera

Science.gov (United States)

Yamashita, Takayuki; Shimamoto, Hiroshi; Funatsu, Ryohei; Mitani, Kohji; Nojiri, Yuji

2006-02-01

We have developed color camera for an 8k x 4k-pixel ultrahigh-definition video system, which is called Super Hi- Vision, with a 5x zoom lens and a signal-processing system incorporating a function for real-time lateral chromatic aberration correction. The chromatic aberration of the lens degrades color image resolution. So in order to develop a compact zoom lens consistent with ultrahigh-resolution characteristics, we incorporated a real-time correction function in the signal-processing system. The signal-processing system has eight memory tables to store the correction data at eight focal length points on the blue and red channels. When the focal length data is inputted from the lens control units, the relevant correction data are interpolated from two of eights correction data tables. This system performs geometrical conversion on both channels using this correction data. This paper describes that the correction function can successfully reduce the lateral chromatic aberration, to an amount small enough to ensure the desired image resolution was achieved over the entire range of the lens in real time.

New insights into the structural organization of eukaryotic and prokaryotic cytoskeletons using cryo-electron tomography

International Nuclear Information System (INIS)

Kuerner, Julia; Medalia, Ohad; Linaroudis, Alexandros A.; Baumeister, Wolfgang

2004-01-01

Cryo-electron tomography (cryo-ET) is an emerging imaging technology that combines the potential of three-dimensional (3-D) imaging at molecular resolution (<5 nm) with a close-to-life preservation of the specimen. In conjunction with pattern recognition techniques, it enables us to map the molecular landscape inside cells. The application of cryo-ET to intact cells provides novel insights into the structure and the spatial organization of the cytoskeleton in prokaryotic and eukaryotic cells

Correcting for color crosstalk and chromatic aberration in multicolor particle shadow velocimetry

International Nuclear Information System (INIS)

McPhail, M J; Fontaine, A A; Krane, M H; Goss, L; Crafton, J

2015-01-01

Color crosstalk and chromatic aberration can bias estimates of fluid velocity measured by color particle shadow velocimetry (CPSV), using multicolor illumination and a color camera. This article describes corrections to remove these bias errors, and their evaluation. Color crosstalk removal is demonstrated with linear unmixing. It is also shown that chromatic aberrations may be removed using either scale calibration, or by processing an image illuminated by all colors simultaneously. CPSV measurements of a fully developed turbulent pipe flow of glycerin were conducted. Corrected velocity statistics from these measurements were compared to both single-color PSV and LDV measurements and showed excellent agreement to fourth-order, to well into the viscous sublayer. Recommendations for practical assessment and correction of color aberration and color crosstalk are discussed. (paper)

Revisiting Cross-Channel Information Transfer for Chromatic Aberration Correction

KAUST Repository

Sun, Tiancheng; Peng, Yifan; Heidrich, Wolfgang

2017-01-01

Image aberrations can cause severe degradation in image quality for consumer-level cameras, especially under the current tendency to reduce the complexity of lens designs in order to shrink the overall size of modules. In simplified optical designs, chromatic aberration can be one of the most significant causes for degraded image quality, and it can be quite difficult to remove in post-processing, since it results in strong blurs in at least some of the color channels. In this work, we revisit the pixel-wise similarity between different color channels of the image and accordingly propose a novel algorithm for correcting chromatic aberration based on this cross-channel correlation. In contrast to recent weak prior-based models, ours uses strong pixel-wise fitting and transfer, which lead to significant quality improvements for large chromatic aberrations. Experimental results on both synthetic and real world images captured by different optical systems demonstrate that the chromatic aberration can be significantly reduced using our approach.

Revisiting Cross-Channel Information Transfer for Chromatic Aberration Correction

KAUST Repository

Sun, Tiancheng

2017-12-25

Image aberrations can cause severe degradation in image quality for consumer-level cameras, especially under the current tendency to reduce the complexity of lens designs in order to shrink the overall size of modules. In simplified optical designs, chromatic aberration can be one of the most significant causes for degraded image quality, and it can be quite difficult to remove in post-processing, since it results in strong blurs in at least some of the color channels. In this work, we revisit the pixel-wise similarity between different color channels of the image and accordingly propose a novel algorithm for correcting chromatic aberration based on this cross-channel correlation. In contrast to recent weak prior-based models, ours uses strong pixel-wise fitting and transfer, which lead to significant quality improvements for large chromatic aberrations. Experimental results on both synthetic and real world images captured by different optical systems demonstrate that the chromatic aberration can be significantly reduced using our approach.

Cryo-electron tomography investigation of serum albumin-camouflaged tobacco mosaic virus nanoparticles.

Science.gov (United States)

Gulati, Neetu M; Pitek, Andrzej S; Steinmetz, Nicole F; Stewart, Phoebe L

2017-03-09

Nanoparticles offer great potential in drug delivery and imaging, but shielding strategies are necessary to increase circulation time and performance. Structure-function studies are required to define the design rules to achieve effective shielding. With several formulations reaching clinical testing and approval, the ability to assess and detail nanoparticle formulations at the single particle level is becoming increasingly important. To address this need, we use cryo-electron tomography (cryo-ET) to investigate stealth-coated nanoparticles. As a model system, we studied the soft matter nanotubes formed by tobacco mosaic virus (TMV) coated with human serum albumin (SA) stealth proteins. Cryo-ET and subtomogram averaging allow for visualization of individual SA molecules and determination of their orientations relative to the TMV surface, and also for measurement of the surface coverage provided by added stealth proteins. This information fills a critical gap in the understanding of the structural morphology of stealth-coated nanoparticles, and therefore cryo-ET may play an important role in guiding the development of future nanoparticle-based therapeutics.

A deep convolutional neural network approach to single-particle recognition in cryo-electron microscopy.

Science.gov (United States)

Zhu, Yanan; Ouyang, Qi; Mao, Youdong

2017-07-21

Single-particle cryo-electron microscopy (cryo-EM) has become a mainstream tool for the structural determination of biological macromolecular complexes. However, high-resolution cryo-EM reconstruction often requires hundreds of thousands of single-particle images. Particle extraction from experimental micrographs thus can be laborious and presents a major practical bottleneck in cryo-EM structural determination. Existing computational methods for particle picking often use low-resolution templates for particle matching, making them susceptible to reference-dependent bias. It is critical to develop a highly efficient template-free method for the automatic recognition of particle images from cryo-EM micrographs. We developed a deep learning-based algorithmic framework, DeepEM, for single-particle recognition from noisy cryo-EM micrographs, enabling automated particle picking, selection and verification in an integrated fashion. The kernel of DeepEM is built upon a convolutional neural network (CNN) composed of eight layers, which can be recursively trained to be highly "knowledgeable". Our approach exhibits an improved performance and accuracy when tested on the standard KLH dataset. Application of DeepEM to several challenging experimental cryo-EM datasets demonstrated its ability to avoid the selection of un-wanted particles and non-particles even when true particles contain fewer features. The DeepEM methodology, derived from a deep CNN, allows automated particle extraction from raw cryo-EM micrographs in the absence of a template. It demonstrates an improved performance, objectivity and accuracy. Application of this novel method is expected to free the labor involved in single-particle verification, significantly improving the efficiency of cryo-EM data processing.

Research on the Phase Aberration Correction with a Deformable Mirror Controlled by a Genetic Algorithm

International Nuclear Information System (INIS)

Yang, P; Hu, S J; Chen, S Q; Yang, W; Xu, B; Jiang, W H

2006-01-01

In order to improve laser beam quality, a real number encoding genetic algorithm based on adaptive optics technology was presented. This algorithm was applied to control a 19-channel deformable mirror to correct phase aberration in laser beam. It is known that when traditional adaptive optics system is used to correct laser beam wave-front phase aberration, a precondition is to measure the phase aberration information in the laser beam. However, using genetic algorithms, there is no necessary to know the phase aberration information in the laser beam beforehand. The only parameter need to know is the Light intensity behind the pinhole on the focal plane. This parameter was used as the fitness function for the genetic algorithm. Simulation results show that the optimal shape of the 19-channel deformable mirror applied to correct the phase aberration can be ascertained. The peak light intensity was improved by a factor of 21, and the encircled energy strehl ratio was increased to 0.34 from 0.02 as the phase aberration was corrected with this technique

Tools for macromolecular model building and refinement into electron cryo-microscopy reconstructions

Energy Technology Data Exchange (ETDEWEB)

Brown, Alan; Long, Fei; Nicholls, Robert A.; Toots, Jaan; Emsley, Paul; Murshudov, Garib, E-mail: garib@mrc-lmb.cam.ac.uk [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH (United Kingdom)

2015-01-01

A description is given of new tools to facilitate model building and refinement into electron cryo-microscopy reconstructions. The recent rapid development of single-particle electron cryo-microscopy (cryo-EM) now allows structures to be solved by this method at resolutions close to 3 Å. Here, a number of tools to facilitate the interpretation of EM reconstructions with stereochemically reasonable all-atom models are described. The BALBES database has been repurposed as a tool for identifying protein folds from density maps. Modifications to Coot, including new Jiggle Fit and morphing tools and improved handling of nucleic acids, enhance its functionality for interpreting EM maps. REFMAC has been modified for optimal fitting of atomic models into EM maps. As external structural information can enhance the reliability of the derived atomic models, stabilize refinement and reduce overfitting, ProSMART has been extended to generate interatomic distance restraints from nucleic acid reference structures, and a new tool, LIBG, has been developed to generate nucleic acid base-pair and parallel-plane restraints. Furthermore, restraint generation has been integrated with visualization and editing in Coot, and these restraints have been applied to both real-space refinement in Coot and reciprocal-space refinement in REFMAC.

Tools for macromolecular model building and refinement into electron cryo-microscopy reconstructions

International Nuclear Information System (INIS)

Brown, Alan; Long, Fei; Nicholls, Robert A.; Toots, Jaan; Emsley, Paul; Murshudov, Garib

2015-01-01

A description is given of new tools to facilitate model building and refinement into electron cryo-microscopy reconstructions. The recent rapid development of single-particle electron cryo-microscopy (cryo-EM) now allows structures to be solved by this method at resolutions close to 3 Å. Here, a number of tools to facilitate the interpretation of EM reconstructions with stereochemically reasonable all-atom models are described. The BALBES database has been repurposed as a tool for identifying protein folds from density maps. Modifications to Coot, including new Jiggle Fit and morphing tools and improved handling of nucleic acids, enhance its functionality for interpreting EM maps. REFMAC has been modified for optimal fitting of atomic models into EM maps. As external structural information can enhance the reliability of the derived atomic models, stabilize refinement and reduce overfitting, ProSMART has been extended to generate interatomic distance restraints from nucleic acid reference structures, and a new tool, LIBG, has been developed to generate nucleic acid base-pair and parallel-plane restraints. Furthermore, restraint generation has been integrated with visualization and editing in Coot, and these restraints have been applied to both real-space refinement in Coot and reciprocal-space refinement in REFMAC

Effects of SMILE and Trans-PRK on corneal higher order aberrations after myopic correction

Directory of Open Access Journals (Sweden)

Jiao Zhao

2018-02-01

Full Text Available AIM:To observe the effects of small incision lenticule extraction(SMILEand trans-epithelial photorefractive keratectomy(Trans-PRKon corneal horizontal coma, vertical coma, and spherical aberration and total higher order aberrations after refractive correction for myopia. METHODS: This was a prospective non-randomized cohort study. The cohort included 40 patients(80 eyeswith myopia, who received refraction correction surgery from December 2016 to February 2017 in Leshan Ophthalmic Center. Twenty patients(40 eyesreceived SMILE surgery and the other 20 patients(40 eyesreceived Trans-PRK surgery. Corneal aberrations were determined by a high-resolution Pentacam Scheimpflug camera before the surgery and at 1 and 3mo after the operation. Statistical analyses were performed using analysis of variance of repeated measures. RESULTS: At 1 and 3mo post-operation, the uncorrected visual acuity in both groups was better than or equal to the preoperative best corrected visual acuity. The preoperative corneal aberrations showed no significant difference between the two groups(P>0.05. Significantly higher aberration was found after the surgery in both groups(PP>0.05. Post-operation, horizontal and vertical coma had no significant difference between the two groups(P>0.05, while SMILE group showed lower spherical aberration and lower total higher order aberration than Trans-PRK group(PCONCLUSION: Both SMILE and Trans-PRK increase corneal aberration and their effects on horizontal and vertical coma are similar. However, SMILE has a minor influence on spherical aberration and total high order aberration than Trans-PRK.

Correction for polychromatic aberration in computed tomography images

International Nuclear Information System (INIS)

Naparstek, A.

1979-01-01

A method and apparatus for correcting a computed tomography image for polychromatic aberration caused by the non-linear interaction (i.e. the energy dependent attenuation characteristics) of different body constituents, such as bone and soft tissue, with a polychromatic X-ray beam are described in detail. An initial image is conventionally computed from path measurements made as source and detector assembly scan a body section. In the improvement, each image element of the initial computed image representing attenuation is recorded in a store and is compared with two thresholds, one representing bone and the other soft tissue. Depending on the element value relative to the thresholds, a proportion of the respective constituent is allocated to that element location and corresponding bone and soft tissue projections are determined and stored. An error projection generator calculates projections of polychromatic aberration errors in the raw image data from recalled bone and tissue projections using a multidimensional polynomial function which approximates the non-linear interaction involved. After filtering, these are supplied to an image reconstruction computer to compute image element correction values which are subtracted from raw image element values to provide a corrected reconstructed image for display. (author)

Fine structure of granal thylakoid membrane organization using cryo electron tomography

NARCIS (Netherlands)

Kouril, Roman; Oostergetel, Gert T.; Boekema, Egbert J.

The architecture of grana membranes from spinach chloroplasts was studied by cryo electron tomography. Tomographic reconstructions of ice-embedded isolated grana stacks enabled to resolve features of photosystem II (PSII) in the native membrane and to assign the absolute orientation of individual

Phasing of the Triatoma virus diffraction data using a cryo-electron microscopy reconstruction

International Nuclear Information System (INIS)

Estrozi, L.F.; Neumann, E.; Squires, G.; Rozas-Dennis, G.; Costabel, M.; Rey, F.A.; Guerin, D.M.A.; Navaza, J.

2008-01-01

The blood-sucking reduviid bug Triatoma infestans, one of the most important vector of American human trypanosomiasis (Chagas disease) is infected by the Triatoma virus (TrV). TrV has been classified as a member of the Cripavirus genus (type cricket paralysis virus) in the Dicistroviridae family. This work presents the three-dimensional cryo-electron microscopy (cryo-EM) reconstruction of the TrV capsid at about 25 A resolution and its use as a template for phasing the available crystallographic data by the molecular replacement method. The main structural differences between the cryo-EM reconstruction of TrV and other two viruses, one from the same family, the cricket paralysis virus (CrPV) and the human rhinovirus 16 from the Picornaviridae family are presented and discussed

Non-common path aberration correction in an adaptive optics scanning ophthalmoscope.

Science.gov (United States)

Sulai, Yusufu N; Dubra, Alfredo

2014-09-01

The correction of non-common path aberrations (NCPAs) between the imaging and wavefront sensing channel in a confocal scanning adaptive optics ophthalmoscope is demonstrated. NCPA correction is achieved by maximizing an image sharpness metric while the confocal detection aperture is temporarily removed, effectively minimizing the monochromatic aberrations in the illumination path of the imaging channel. Comparison of NCPA estimated using zonal and modal orthogonal wavefront corrector bases provided wavefronts that differ by ~λ/20 in root-mean-squared (~λ/30 standard deviation). Sequential insertion of a cylindrical lens in the illumination and light collection paths of the imaging channel was used to compare image resolution after changing the wavefront correction to maximize image sharpness and intensity metrics. Finally, the NCPA correction was incorporated into the closed-loop adaptive optics control by biasing the wavefront sensor signals without reducing its bandwidth.

Structural defects in cubic semiconductors characterized by aberration-corrected scanning transmission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Arroyo Rojas Dasilva, Yadira; Kozak, Roksolana; Erni, Rolf; Rossell, Marta D., E-mail: marta.rossell@empa.ch

2017-05-15

The development of new electro-optical devices and the realization of novel types of transistors require a profound understanding of the structural characteristics of new semiconductor heterostructures. This article provides a concise review about structural defects which occur in semiconductor heterostructures on the basis of micro-patterned Si substrates. In particular, one- and two-dimensional crystal defects are being discussed which are due to the plastic relaxation of epitaxial strain caused by the misfit of crystal lattices. Besides a few selected examples from literature, we treat in particular crystal defects occurring in GaAs/Si, Ge/Si and β-SiC/Si structures which are studied by high-resolution annular dark-field scanning transmission electron microscopy. The relevance of this article is twofold; firstly, it should provide a collection of data which are of help for the identification and characterization of defects in cubic semiconductors by means of atomic-resolution imaging, and secondly, the experimental data shall provide a basis for advancing the understanding of device characteristics with the aid of theoretical modelling by considering the defective nature of strained semiconductor heterostructures. - Highlights: • The heterogeneous integration of high-quality compound semiconductors remains a challenge. • Lattice defects cause severe degradation of the semiconductor device performances. • Aberration-corrected HAADF-STEM allows atomic-scale characterization of defects. • An overview of lattice defects found in cubic semiconductors is presented. • Theoretical modelling and calculations are needed to determine the defect properties.

Cryo-transmission electron tomography of native casein micelles from bovine milk

Science.gov (United States)

Trejo, R.; Dokland, T.; Jurat-Fuentes, J.; Harte, F.

2013-01-01

Caseins are the principal protein components in milk and an important ingredient in the food industry. In liquid milk, caseins are found as micelles of casein proteins and colloidal calcium nanoclusters. Casein micelles were isolated from raw skim milk by size exclusion chromatography and suspended in milk protein-free serum produced by ultrafiltration (molecular weight cut-off of 3 kDa) of raw skim milk. The micelles were imaged by cryo-electron microscopy and subjected to tomographic reconstruction methods to visualize the 3-dimensional and internal organization of native casein micelles. This provided new insights into the internal architecture of the casein micelle that had not been apparent from prior cryo-transmission electron microscopy studies. This analysis demonstrated the presence of water-filled cavities (~20 to 30 nm in diameter), channels (diameter greater than ~5 nm), and several hundred high-density nanoclusters (6 to 12 nm in diameter) within the interior of the micelles. No spherical protein submicellar structures were observed. PMID:22118067

Integral image rendering procedure for aberration correction and size measurement.

Science.gov (United States)

Sommer, Holger; Ihrig, Andreas; Ebenau, Melanie; Flühs, Dirk; Spaan, Bernhard; Eichmann, Marion

2014-05-20

The challenge in rendering integral images is to use as much information preserved by the light field as possible to reconstruct a captured scene in a three-dimensional way. We propose a rendering algorithm based on the projection of rays through a detailed simulation of the optical path, considering all the physical properties and locations of the optical elements. The rendered images contain information about the correct size of imaged objects without the need to calibrate the imaging device. Additionally, aberrations of the optical system may be corrected, depending on the setup of the integral imaging device. We show simulation data that illustrates the aberration correction ability and experimental data from our plenoptic camera, which illustrates the capability of our proposed algorithm to measure size and distance. We believe this rendering procedure will be useful in the future for three-dimensional ophthalmic imaging of the human retina.

Development of superconducting cryo-electron microscope and its applications

International Nuclear Information System (INIS)

Iwatsuki, Masashi

1988-01-01

Recently, a superconducting cryo-electron microscope in which specimens are cooled to the liquid helium temperature (4.2 K) has been developed. The main components and functional features of this new microscope are reported together with application data on polyethylene, poly (4-methyl-1-pentene), valonia cellulose, rock salt, ice crystallites and ceramic superconductor. The resistance to electron radiation damage, of beam-sensitive specimens including polymers has been increased more than ten times. Thus, the microscope has made it possible to take high resolution images and to analyze the crystal-structure of micro-areas. (orig.) [de

Pleomorphism and Viability of the Lyme Disease Pathogen Borrelia burgdorferi Exposed to Physiological Stress Conditions: A Correlative Cryo-Fluorescence and Cryo-Scanning Electron Microscopy Study

Czech Academy of Sciences Publication Activity Database

Vancová, Marie; Rudenko, Natalia; Vaněček, Jiří; Golovchenko, Maryna; Strnad, Martin; Rego, Ryan O. M.; Tichá, Lucie; Grubhoffer, Libor; Nebesářová, Jana

2017-01-01

Roč. 8, 11 April (2017), č. článku 596. ISSN 1664-302X R&D Projects: GA TA ČR(CZ) TE01020118; GA MŠk(CZ) LM2015062 EU Projects: European Commission(XE) 278976 - ANTIGONE Institutional support: RVO:60077344 Keywords : cryo-fluorescence * cryo-scanning electron microscopy * Borrelia burgdorferi * Lyme disease * round body * pleomorphism * viability staining Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: 2.11 Other engineering and technologies Impact factor: 4.076, year: 2016

Quantum ballistic transistor and low noise HEMT for cryo-electronics lower than 4.2 K

International Nuclear Information System (INIS)

Gremion, E.

2008-01-01

Next generations of cryo-detectors, widely used in physics of particles and physics of universe, will need in the future high-performance cryo-electronics less noisy and closer to the detector. Within this context, this work investigates properties of two dimensional electron gas GaAlAs/GaAs by studying two components, quantum point contact (QPC) and high electron mobility transistor (HEMT). Thanks to quantized conductance steps in QPC, we have realized a quantum ballistic transistor (voltage gain higher than 1), a new component useful for cryo-electronics thanks to its operating temperature and weak power consumption (about 1 nW). Moreover, the very low capacity of this component leads to promising performances for multiplexing low temperature bolometer dedicated to millimetric astronomy. The second study focused on HEMT with very high quality 2DEG. At 4.2 K, a voltage gain higher than 20 can be obtained with a very low power dissipation of less than 100 μW. Under the above experimental conditions, an equivalent input voltage noise of 1.2 nV/√(Hz) at 1 kHz and 0.12 nV/√(Hz) at 100 kHz has been reached. According to the Hooge formula, these noise performances are get by increasing gate capacity estimated to 60 pF. (author)

Ultrashort echo-time MRI versus CT for skull aberration correction in MR-guided transcranial focused ultrasound: In vitro comparison on human calvaria.

Science.gov (United States)

Miller, G Wilson; Eames, Matthew; Snell, John; Aubry, Jean-François

2015-05-01

Transcranial magnetic resonance-guided focused ultrasound (TcMRgFUS) brain treatment systems compensate for skull-induced beam aberrations by adjusting the phase and amplitude of individual ultrasound transducer elements. These corrections are currently calculated based on a preacquired computed tomography (CT) scan of the patient's head. The purpose of the work presented here is to demonstrate the feasibility of using ultrashort echo-time magnetic resonance imaging (UTE MRI) instead of CT to calculate and apply aberration corrections on a clinical TcMRgFUS system. Phantom experiments were performed in three ex-vivo human skulls filled with tissue-mimicking hydrogel. Each skull phantom was imaged with both CT and UTE MRI. The MR images were then segmented into "skull" and "not-skull" pixels using a computationally efficient, threshold-based algorithm, and the resulting 3D binary skull map was converted into a series of 2D virtual CT images. Each skull was mounted in the head transducer of a clinical TcMRgFUS system (ExAblate Neuro, Insightec, Israel), and transcranial sonications were performed using a power setting of approximately 750 acoustic watts at several different target locations within the electronic steering range of the transducer. Each target location was sonicated three times: once using aberration corrections calculated from the actual CT scan, once using corrections calculated from the MRI-derived virtual CT scan, and once without applying any aberration correction. MR thermometry was performed in conjunction with each 10-s sonication, and the highest single-pixel temperature rise and surrounding-pixel mean were recorded for each sonication. The measured temperature rises were ∼ 45% larger for aberration-corrected sonications than for noncorrected sonications. This improvement was highly significant (p skull-induced ultrasound aberration corrections. Their results suggest that UTE MRI could be used instead of CT to implement such corrections on

Cryo-transmission electron microscopy of Ag nanoparticles grown on an ionic liquid substrate

KAUST Repository

Anjum, Dalaver H.

2010-07-01

We report a novel method of growing silver nanostructures by cathodic sputtering onto an ionic liquid (IL) and our visualization by transmission cryo-electron microscopy to avoid beam-induced motion of the nanoparticles. By freezing the IL suspension and controlling electron dose, we can assess properties of particle size, morphology, crystallinity, and aggregation in situ and at high detail. We observed round silver nanoparticles with a well-defined diameter of 7.0 ± 1.5 nm that are faceted with crystalline cubic structures and ∼80% of the particles have multiply twinned faults. We also applied cryo-electron tomography to investigate the structure of the nanoparticles and to directly visualize the IL wetting around them. In addition to particles, we observed nanorods that appear to have assembled from individual nanoparticles. Reexamination of the samples after 4-5 days from initial preparation showed significant changes in morphology, and potential mechanisms for this are discussed. © 2010 Materials Research Society.

Cryo-transmission electron microscopy of Ag nanoparticles grown on an ionic liquid substrate

KAUST Repository

Anjum, Dalaver H.; Stiger, Rebecca M.; Finley, James J.; Conway, James F.

2010-01-01

We report a novel method of growing silver nanostructures by cathodic sputtering onto an ionic liquid (IL) and our visualization by transmission cryo-electron microscopy to avoid beam-induced motion of the nanoparticles. By freezing the IL

Atomic resolution imaging of YAlO{sub 3}: Ce in the chromatic and spherical aberration corrected PICO electron microscope

Energy Technology Data Exchange (ETDEWEB)

Jin, Lei [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Jülich-Aachen Research Alliance (JARA), Forschungszentrum Jülich GmbH, 52425 Jülich (Germany); Barthel, Juri [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Jülich-Aachen Research Alliance (JARA), Forschungszentrum Jülich GmbH, 52425 Jülich (Germany); Central Facility for Electron Microscopy, RWTH Aachen University, 52074 Aachen (Germany); Jia, Chun-Lin [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Jülich-Aachen Research Alliance (JARA), Forschungszentrum Jülich GmbH, 52425 Jülich (Germany); School of Electronic and Information Engineering and State Key Laboratory for Mechanical Behaviour of Materials, Xi' an Jiaotong University, Xi' an 710049 (China); Urban, Knut W., E-mail: k.urban@fz-juelich.de [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Jülich-Aachen Research Alliance (JARA), Forschungszentrum Jülich GmbH, 52425 Jülich, (Germany); School of Electronic and Information Engineering and State Key Laboratory for Mechanical Behaviour of Materials, Xi' an Jiaotong University, Xi' an 710049 (China)

2017-05-15

Highlights: • First time resolution of 57 pm atom separations by HRTEM with 200 keV electrons. • Quantification of the image spread by absolute matching of experiment and simulation. • An information limit of 52 pm is deduced from the determined image spread. • Substantial deviations from the bulk structure are observed for the ultra-thin sample. - Abstract: The application of combined chromatic and spherical aberration correction in high-resolution transmission electron microscopy enables a significant improvement of the spatial resolution down to 50 pm. We demonstrate that such a resolution can be achieved in practice at 200 kV. Diffractograms of images of gold nanoparticles on amorphous carbon demonstrate corresponding information transfer. The Y atom pairs in [010] oriented yttrium orthoaluminate are successfully imaged together with the Al and the O atoms. Although the 57 pm pair separation is well demonstrated separations between 55 pm and 80 pm are measured. This observation is tentatively attributed to structural relaxations and surface reconstruction in the very thin samples used. Quantification of the resolution limiting effective image spread is achieved based on an absolute match between experimental and simulated image intensity distributions.

Aberration-corrected scanning transmission electron microscopy for complex transition metal oxides

Science.gov (United States)

Qing-Hua, Zhang; Dong-Dong, Xiao; Lin, Gu

2016-06-01

Lattice, charge, orbital, and spin are the four fundamental degrees of freedom in condensed matter, of which the interactive coupling derives tremendous novel physical phenomena, such as high-temperature superconductivity (high-T c SC) and colossal magnetoresistance (CMR) in strongly correlated electronic system. Direct experimental observation of these freedoms is essential to understanding the structure-property relationship and the physics behind it, and also indispensable for designing new materials and devices. Scanning transmission electron microscopy (STEM) integrating multiple techniques of structure imaging and spectrum analysis, is a comprehensive platform for providing structural, chemical and electronic information of materials with a high spatial resolution. Benefiting from the development of aberration correctors, STEM has taken a big breakthrough towards sub-angstrom resolution in last decade and always steps forward to improve the capability of material characterization; many improvements have been achieved in recent years, thereby giving an in-depth insight into material research. Here, we present a brief review of the recent advances of STEM by some representative examples of perovskite transition metal oxides; atomic-scale mapping of ferroelectric polarization, octahedral distortions and rotations, valence state, coordination and spin ordering are presented. We expect that this brief introduction about the current capability of STEM could facilitate the understanding of the relationship between functional properties and these fundamental degrees of freedom in complex oxides. Project supported by the National Key Basic Research Project, China (Grant No. 2014CB921002), the Strategic Priority Research Program of Chinese Academy of Sciences (Grant No. XDB07030200), and the National Natural Science Foundation of China (Grant Nos. 51522212 and 51421002).

Spherical aberration correction with an in-lens N-fold symmetric line currents model.

Science.gov (United States)

Hoque, Shahedul; Ito, Hiroyuki; Nishi, Ryuji

2018-04-01

In our previous works, we have proposed N-SYLC (N-fold symmetric line currents) models for aberration correction. In this paper, we propose "in-lens N-SYLC" model, where N-SYLC overlaps rotationally symmetric lens. Such overlap is possible because N-SYLC is free of magnetic materials. We analytically prove that, if certain parameters of the model are optimized, an in-lens 3-SYLC (N = 3) doublet can correct 3rd order spherical aberration. By computer simulation, we show that the required excitation current for correction is less than 0.25 AT for beam energy 5 keV, and the beam size after correction is smaller than 1 nm at the corrector image plane for initial slope less than 4 mrad. Copyright © 2018 Elsevier B.V. All rights reserved.

High-resolution structure of the Shigella type-III secretion needle by solid-state NMR and cryo-electron microscopy

Science.gov (United States)

Demers, Jean-Philippe; Habenstein, Birgit; Loquet, Antoine; Kumar Vasa, Suresh; Giller, Karin; Becker, Stefan; Baker, David; Lange, Adam; Sgourakis, Nikolaos G.

2014-09-01

We introduce a general hybrid approach for determining the structures of supramolecular assemblies. Cryo-electron microscopy (cryo-EM) data define the overall envelope of the assembly and rigid-body orientation of the subunits while solid-state nuclear magnetic resonance (ssNMR) chemical shifts and distance constraints define the local secondary structure, protein fold and inter-subunit interactions. Finally, Rosetta structure calculations provide a general framework to integrate the different sources of structural information. Combining a 7.7-Å cryo-EM density map and 996 ssNMR distance constraints, the structure of the type-III secretion system needle of Shigella flexneri is determined to a precision of 0.4 Å. The calculated structures are cross-validated using an independent data set of 691 ssNMR constraints and scanning transmission electron microscopy measurements. The hybrid model resolves the conformation of the non-conserved N terminus, which occupies a protrusion in the cryo-EM density, and reveals conserved pore residues forming a continuous pattern of electrostatic interactions, thereby suggesting a mechanism for effector protein translocation.

Chromatic aberrations correction for imaging spectrometer based on acousto-optic tunable filter with two transducers.

Science.gov (United States)

Zhao, Huijie; Wang, Ziye; Jia, Guorui; Zhang, Ying; Xu, Zefu

2017-10-02

The acousto-optic tunable filter (AOTF) with wide wavelength range and high spectral resolution has long crystal and two transducers. A longer crystal length leads to a bigger chromatic focal shift and the double-transducer arrangement induces angular mutation in diffracted beam, which increase difficulty in longitudinal and lateral chromatic aberration correction respectively. In this study, the two chromatic aberrations are analyzed quantitatively based on an AOTF optical model and a novel catadioptric dual-path configuration is proposed to correct both the chromatic aberrations. The test results exhibit effectiveness of the optical configuration for this type of AOTF-based imaging spectrometer.

Investigation of focusing and correcting aberrations with binary amplitude and polarization modulation.

Science.gov (United States)

Fiala, Peter; Li, Yunqi; Dorrer, Christophe

2018-02-01

We investigate the focusing and correcting wavefront aberration of an optical wave using binary amplitude and polarization modulation. Focusing is performed by selectively modulating the field in different zones of the pupil to obtain on-axis constructive interference at a given distance. The conventional Soret zone plate (binary amplitude profile) is expanded to a polarization Soret zone plate with twice the focusing efficiency. Binary pixelated devices that approximate the sinusoidal transmission profile of a Gabor zone plate by spatial dithering are also investigated with amplitude and polarization modulation. Wavefront aberrations are corrected by modulation of the field in the pupil plane to prevent destructive interference in the focal plane of an ideal focusing element. Polarization modulation improves the efficiency obtained by amplitude-only modulation, with a gain that depends on the aberration. Experimental results obtained with Cr-on-glass devices for amplitude modulation and liquid crystal devices operating in the Mauguin condition for polarization modulation are in very good agreement with simulations.

Single particle and molecular assembly analysis of polyribosomes by single- and double-tilt cryo electron tomography

Energy Technology Data Exchange (ETDEWEB)

Myasnikov, Alexander G. [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Department of Integrative Structural Biology, Centre National de la Recherche Scientifique (CNRS) UMR 7104/ Institut National de la Santé de la Recherche Médicale INSERM U964/ Université de Strasbourg, 1 rue Laurent Fries, 67404 Illkirch (France); Afonina, Zhanna A. [Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region (Russian Federation); Klaholz, Bruno P., E-mail: klaholz@igbmc.fr [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Department of Integrative Structural Biology, Centre National de la Recherche Scientifique (CNRS) UMR 7104/ Institut National de la Santé de la Recherche Médicale INSERM U964/ Université de Strasbourg, 1 rue Laurent Fries, 67404 Illkirch (France)

2013-03-15

Cryo electron tomography (cryo-ET) can provide cellular and molecular structural information on various biological samples. However, the detailed interpretation of tomograms reconstructed from single-tilt data tends to suffer from low signal-to-noise ratio and artefacts caused by some systematically missing angular views. While these can be overcome by sub-tomogram averaging, they remain limiting for the analysis of unique structures. Double-tilt ET can improve the tomogram quality by acquiring a second tilt series after an in-plane rotation, but its usage is not widespread yet because it is considered technically demanding and it is rarely used under cryo conditions. Here we show that double-tilt cryo-ET improves the quality of 3D reconstructions so significantly that even single particle analysis can be envisaged despite of the intrinsically low image contrast obtained from frozen-hydrated specimens. This is illustrated by the analysis of eukaryotic polyribosomes in which individual ribosomes were reconstructed using single-tilt, partial and full double-tilt geometries. The improved tomograms favour the faster convergence of iterative sub-tomogram averaging and allow a better 3D classification using multivariate statistical analysis. Our study of single particles and molecular assemblies within polysomes illustrates that the dual-axis approach is particularly useful for cryo applications of ET, both for unique objects and for structures that can be classified and averaged. - Highlights: ► Double-tilt cryo-ET improves 3D reconstructions thus making single particle analysis possible. ► Dual-axis cryo-ET data favour a faster convergence of iterative sub-tomogram averaging. ► Individual ribosomes were reconstructed from single-tilt, partial/ full double-tilt geometries. ► Double-tilt cryo-ET facilitates analysis of larger molecular assemblies such as in cell sections. ► Dual-axis cryo-ET is applicable to unique objects and to structures that can be

Simultaneous correction of large low-order and high-order aberrations with a new deformable mirror technology

Science.gov (United States)

Rooms, F.; Camet, S.; Curis, J. F.

2010-02-01

A new technology of deformable mirror will be presented. Based on magnetic actuators, these deformable mirrors feature record strokes (more than +/- 45μm of astigmatism and focus correction) with an optimized temporal behavior. Furthermore, the development has been made in order to have a large density of actuators within a small clear aperture (typically 52 actuators within a diameter of 9.0mm). We will present the key benefits of this technology for vision science: simultaneous correction of low and high order aberrations, AO-SLO image without artifacts due to the membrane vibration, optimized control, etc. Using recent papers published by Doble, Thibos and Miller, we show the performances that can be achieved by various configurations using statistical approach. The typical distribution of wavefront aberrations (both the low order aberration (LOA) and high order aberration (HOA)) have been computed and the correction applied by the mirror. We compare two configurations of deformable mirrors (52 and 97 actuators) and highlight the influence of the number of actuators on the fitting error, the photon noise error and the effective bandwidth of correction.

Correction of 157-nm lens based on phase ring aberration extraction method

Science.gov (United States)

Meute, Jeff; Rich, Georgia K.; Conley, Will; Smith, Bruce W.; Zavyalova, Lena V.; Cashmore, Julian S.; Ashworth, Dominic; Webb, James E.; Rich, Lisa

2004-05-01

Early manufacture and use of 157nm high NA lenses has presented significant challenges including: intrinsic birefringence correction, control of optical surface contamination, and the use of relatively unproven materials, coatings, and metrology. Many of these issues were addressed during the manufacture and use of International SEMATECH"s 0.85NA lens. Most significantly, we were the first to employ 157nm phase measurement interferometry (PMI) and birefringence modeling software for lens optimization. These efforts yielded significant wavefront improvement and produced one of the best wavefront-corrected 157nm lenses to date. After applying the best practices to the manufacture of the lens, we still had to overcome the difficulties of integrating the lens into the tool platform at International SEMATECH instead of at the supplier facility. After lens integration, alignment, and field optimization were complete, conventional lithography and phase ring aberration extraction techniques were used to characterize system performance. These techniques suggested a wavefront error of approximately 0.05 waves RMS--much larger than the 0.03 waves RMS predicted by 157nm PMI. In-situ wavefront correction was planned for in the early stages of this project to mitigate risks introduced by the use of development materials and techniques and field integration of the lens. In this publication, we document the development and use of a phase ring aberration extraction method for characterizing imaging performance and a technique for correcting aberrations with the addition of an optical compensation plate. Imaging results before and after the lens correction are presented and differences between actual and predicted results are discussed.

Automated aberration correction of arbitrary laser modes in high numerical aperture systems

OpenAIRE

Hering, Julian; Waller, Erik H.; Freymann, Georg von

2016-01-01

Controlling the point-spread-function in three-dimensional laser lithography is crucial for fabricating structures with highest definition and resolution. In contrast to microscopy, aberrations have to be physically corrected prior to writing, to create well defined doughnut modes, bottlebeams or multi foci modes. We report on a modified Gerchberg-Saxton algorithm for spatial-light-modulator based automated aberration compensation to optimize arbitrary laser-modes in a high numerical aperture...

Automated aberration correction of arbitrary laser modes in high numerical aperture systems.

Science.gov (United States)

Hering, Julian; Waller, Erik H; Von Freymann, Georg

2016-12-12

Controlling the point-spread-function in three-dimensional laser lithography is crucial for fabricating structures with highest definition and resolution. In contrast to microscopy, aberrations have to be physically corrected prior to writing, to create well defined doughnut modes, bottlebeams or multi foci modes. We report on a modified Gerchberg-Saxton algorithm for spatial-light-modulator based automated aberration compensation to optimize arbitrary laser-modes in a high numerical aperture system. Using circularly polarized light for the measurement and first-guess initial conditions for amplitude and phase of the pupil function our scalar approach outperforms recent algorithms with vectorial corrections. Besides laser lithography also applications like optical tweezers and microscopy might benefit from the method presented.

Preservation of high resolution protein structure by cryo-electron microscopy of vitreous sections

International Nuclear Information System (INIS)

Sader, Kasim; Studer, Daniel; Zuber, Benoit; Gnaegi, Helmut; Trinick, John

2009-01-01

We have quantitated the degree of structural preservation in cryo-sections of a vitrified biological specimen. Previous studies have used sections of periodic specimens to assess the resolution present, but preservation before sectioning was not assessed and so the damage due particularly to cutting was not clear. In this study large single crystals of lysozyme were vitrified and from these X-ray diffraction patterns extending to better than 2.1 A were obtained. The crystals were high pressure frozen in 30% dextran, and cryo-sectioned using a diamond knife. In the best case, preservation to a resolution of 7.9 A was shown by electron diffraction, the first observation of sub-nanometre structural preservation in a vitreous section.

Direct observation of dislocation dissociation and Suzuki segregation in a Mg–Zn–Y alloy by aberration-corrected scanning transmission electron microscopy

International Nuclear Information System (INIS)

Yang Zhiqing; Chisholm, Matthew F.; Duscher, Gerd; Ma Xiuliang; Pennycook, Stephen J.

2013-01-01

Crystal defects in a plastically deformed Mg–Zn–Y alloy have been studied on the atomic scale using aberration-corrected scanning transmission electron microscopy, providing important structural data for understanding the material’s deformation behavior and strengthening mechanisms. Atomic scale structures of deformation stacking faults resulting from dissociation of different types of dislocations have been characterized experimentally, and modeled. Suzuki segregation of Zn and Y along stacking faults formed through dislocation dissociation during plastic deformation at 300 °C is confirmed experimentally on the atomic level. The stacking fault energy of the Mg–Zn–Y alloy is evaluated to be in the range of 4.0–10.3 mJ m −2 . The newly formed nanometer-wide stacking faults with their Zn/Y segregation in Mg grains play an important role in the superior strength of this alloy at elevated temperatures.

Coherence and diffraction limited resolution in microscopic OCT by a unified approach for the correction of dispersion and aberrations

Science.gov (United States)

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.

Never at rest: insights into the conformational dynamics of ion channels from cryo-electron microscopy.

Science.gov (United States)

Lau, Carus; Hunter, Mark J; Stewart, Alastair; Perozo, Eduardo; Vandenberg, Jamie I

2018-04-01

The tightly regulated opening and closure of ion channels underlies the electrical signals that are vital for a wide range of physiological processes. Two decades ago the first atomic level view of ion channel structures led to a detailed understanding of ion selectivity and conduction. In recent years, spectacular developments in the field of cryo-electron microscopy have resulted in cryo-EM superseding crystallography as the technique of choice for determining near-atomic resolution structures of ion channels. Here, we will review the recent developments in cryo-EM and its specific application to the study of ion channel gating. We will highlight the advantages and disadvantages of the current technology and where the field is likely to head in the next few years. © 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.

Value of lymphocyte cryo-preservation after a radiological or nuclear accident

International Nuclear Information System (INIS)

Laroche, P.; Lataillade, J.J.; Chambrette, V.; Voisin, Ph.

1997-01-01

The conventional cytogenetic method in biological dosimetry is most useful for the estimation of the received radiation dose. It shows resulting unstable chromosomal aberrations (dicentrics, centric rings and fragments) in peripheral blood lymphocytes. This method has been used over the past 30 years and is used in forensic medicine. Nevertheless, it is long and fastidious. Accordingly, the number of simultaneous analyses of blood samples is limited and depends on the capacity of specialized laboratories. This capacity may be insufficient in the case of large scale radiological or nuclear accidents. Cryo-preservation is the usual method to store cells before analysis or use, for instance for biological dosimetry purposes. Some investigations have shown that thawing following freezing may induce cell injury but few studies have been made on the effect of cryo-preservation on cells containing radiation-induced unstable chromosomal aberrations. In this work, lymphocytes were irradiated with 1 to 4 Gy gamma rays and stored in liquid nitrogen. The dicentric and centric ring yields were analysed after storage periods of 1 week, 1 month, 3 months, and 1 year. No difference in aberration frequency from control, unfrozen samples was observed over this period. Lymphocytes stored at -196 deg C for up to least 1 year may therefore be used for chromosome aberration scoring when overexposure to ionizing radiation is suspected. (author)

Depth-resolution imaging of crystalline nanoclusters attached on and embedded in amorphous films using aberration-corrected TEM

Energy Technology Data Exchange (ETDEWEB)

Yamasaki, Jun, E-mail: yamasaki@uhvem.osaka-u.ac.jp [Research Center for Ultra-High Voltage Electron Microscopy, Osaka University, 7-1 Mihogaoka, Ibaraki, Osaka 567-0047 (Japan); EcoTopia Science Institute, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan); Mori, Masayuki [Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan); Hirata, Akihiko [Advanced Institute for Materials Research, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577 (Japan); Hirotsu, Yoshihiko [Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047 (Japan); Tanaka, Nobuo [EcoTopia Science Institute, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603 (Japan)

2015-04-15

For observations of crystalline nanoclusters, the features and capabilities of depth-resolution imaging by aberration-corrected transmission electron microscopy (TEM) were investigated using image simulations and experiments for two types of samples. The first sample was gold clusters attached on an amorphous carbon film. The experimental through-focal series indicated that the focal plane for the cluster was shifted 3 nm from that for the supporting film. This difference is due to the depth-resolution imaging of the cluster and film, the mid-planes of which are separated by 3 nm along the depth direction (the electron incident direction). On the basis of this information, the three-dimensional configuration of the sample, such as the film thickness of 2 nm, was successfully illustrated. The second sample was a Zr{sub 66.7}Ni{sub 33.3} metallic glass including a medium-range-order (MRO) structure, which was approximately considered to be a crystalline cluster with a diameter of 1.6 nm. In the experimental through-focal series, the lattice fringe of the MRO cluster was visible at limited focal conditions. Image simulations reproduced well the focal conditions and also indicated a structural condition for the visualization that the embedded cluster must be apart from the mid-plane of the matrix film. Similar to the case of the first sample, this result can be explained by the idea that the “effective focal planes” for the film and cluster are at different heights. This type of depth-resolution phase contrast imaging is possible only in aberration-corrected TEM and when the sample has a simple structure and is sufficiently thin for the kinematical scattering approximation. - Highlights: • Depth-resolution imaging by aberration-corrected TEM was demonstrated. • Thickness of a carbon film supporting gold nano-crystals was successfully estimated. • A crystalline nanocluster embedded in an amorphous matrix was successfully observed. • It was clarified that

Outline of an ultracorrector compensating for all primary chromatic and geometrical aberrations of charged-particle lenses

International Nuclear Information System (INIS)

Rose, Harald

2004-01-01

A novel ultracorrector is outlined which compensates for the primary and secondary first-order chromatic aberrations and all third-order geometrical aberrations of electron optical systems with a straight axis. Owing to the imposed symmetry conditions on the fields and the paraxial fundamental rays, the corrector does not introduce aberrations with 2-fold symmetry. The chromatic aberrations are corrected by means of crossed electric magnetic quadrupoles while the third-order geometrical aberrations are eliminated by octopoles. By placing these elements at distinct positions within the corrector, it is possible to successively eliminate the third-order aberrations in such a way that each subsequent correction does not affect the aberrations corrected in the preceding correction steps

Practical aspects of Boersch phase contrast electron microscopy of biological specimens

Energy Technology Data Exchange (ETDEWEB)

Walter, Andreas [Max-Planck-Institute of Biophysics, Department of Structural Biology, Max-von-Laue-Str. 3, D-60439 Frankfurt (Germany); Muzik, Heiko; Vieker, Henning; Turchanin, Andrey; Beyer, Andre; Goelzhaeuser, Armin [University of Bielefeld, Physics of Supramolecular Systems and Surfaces, Universitaetsstr. 25, D-33615 Bielefeld (Germany); Lacher, Manfred; Steltenkamp, Siegfried; Schmitz, Sam; Holik, Peter [Caesar Research Center, Ludwig-Erhard-Allee 2, D-53175 Bonn (Germany); Kuehlbrandt, Werner [Max-Planck-Institute of Biophysics, Department of Structural Biology, Max-von-Laue-Str. 3, D-60439 Frankfurt (Germany); Rhinow, Daniel, E-mail: daniel.rhinow@biophys.mpg.de [Max-Planck-Institute of Biophysics, Department of Structural Biology, Max-von-Laue-Str. 3, D-60439 Frankfurt (Germany)

2012-05-15

Implementation of physical phase plates into transmission electron microscopes to achieve in-focus contrast for ice-embedded biological specimens poses several technological challenges. During the last decade several phase plates designs have been introduced and tested for electron cryo-microscopy (cryoEM), including thin film (Zernike) phase plates and electrostatic devices. Boersch phase plates (BPPs) are electrostatic einzel lenses shifting the phase of the unscattered beam by an arbitrary angle. Adjusting the phase shift to 90 Degree-Sign achieves the maximum contrast transfer for phase objects such as biomolecules. Recently, we reported the implementation of a BPP into a dedicated phase contrast aberration-corrected electron microscope (PACEM) and demonstrated its use to generate in-focus contrast of frozen-hydrated specimens. However, a number of obstacles need to be overcome before BPPs can be used routinely, mostly related to the phase plate devices themselves. CryoEM with a physical phase plate is affected by electrostatic charging, obliteration of low spatial frequencies, and mechanical drift. Furthermore, BPPs introduce single sideband contrast (SSB), due to the obstruction of Friedel mates in the diffraction pattern. In this study we address the technical obstacles in detail and show how they may be overcome. We use X-ray photoelectron spectroscopy (XPS) and Auger electron spectroscopy (AES) to identify contaminants responsible for electrostatic charging, which occurs with most phase plates. We demonstrate that obstruction of low-resolution features is significantly reduced by lowering the acceleration voltage of the microscope. Finally, we present computational approaches to correct BPP images for SSB contrast and to compensate for mechanical drift of the BPP. -- Highlights: Black-Right-Pointing-Pointer Various obstacles need to be overcome before Boersch phase plates can be used routinely. Black-Right-Pointing-Pointer Technical problems include

Practical aspects of Boersch phase contrast electron microscopy of biological specimens

International Nuclear Information System (INIS)

Walter, Andreas; Muzik, Heiko; Vieker, Henning; Turchanin, Andrey; Beyer, André; Gölzhäuser, Armin; Lacher, Manfred; Steltenkamp, Siegfried; Schmitz, Sam; Holik, Peter; Kühlbrandt, Werner; Rhinow, Daniel

2012-01-01

Implementation of physical phase plates into transmission electron microscopes to achieve in-focus contrast for ice-embedded biological specimens poses several technological challenges. During the last decade several phase plates designs have been introduced and tested for electron cryo-microscopy (cryoEM), including thin film (Zernike) phase plates and electrostatic devices. Boersch phase plates (BPPs) are electrostatic einzel lenses shifting the phase of the unscattered beam by an arbitrary angle. Adjusting the phase shift to 90° achieves the maximum contrast transfer for phase objects such as biomolecules. Recently, we reported the implementation of a BPP into a dedicated phase contrast aberration-corrected electron microscope (PACEM) and demonstrated its use to generate in-focus contrast of frozen–hydrated specimens. However, a number of obstacles need to be overcome before BPPs can be used routinely, mostly related to the phase plate devices themselves. CryoEM with a physical phase plate is affected by electrostatic charging, obliteration of low spatial frequencies, and mechanical drift. Furthermore, BPPs introduce single sideband contrast (SSB), due to the obstruction of Friedel mates in the diffraction pattern. In this study we address the technical obstacles in detail and show how they may be overcome. We use X-ray photoelectron spectroscopy (XPS) and Auger electron spectroscopy (AES) to identify contaminants responsible for electrostatic charging, which occurs with most phase plates. We demonstrate that obstruction of low-resolution features is significantly reduced by lowering the acceleration voltage of the microscope. Finally, we present computational approaches to correct BPP images for SSB contrast and to compensate for mechanical drift of the BPP. -- Highlights: ► Various obstacles need to be overcome before Boersch phase plates can be used routinely. ► Technical problems include electrostatic charging, mechanical drift, and image artefacts. �

The influence of Cs/Cc correction in analytical imaging and spectroscopy in scanning and transmission electron microscopy

International Nuclear Information System (INIS)

Zaluzec, Nestor J.

2015-01-01

Aberration correction in scanning/transmission electron microscopy (S/TEM) owes much to the efforts of a small dedicated group of innovators. Leading that frontier has been Prof. Harald Rose. To date his leadership and dynamic personality has spearheaded our ability to leave behind many of the limitations imposed by spherical aberration (C s ) in high resolution phase contrast imaging. Following shortly behind, has been the development of chromatic aberration correction (C c ) which augments those accomplishments. In this paper we will review and summarize how the combination of C s /C c technology enhances our ability to conduct hyperspectral imaging and spectroscopy in today's and future computationally mediated experiments in both thin as well as realistic specimens in vacuo and during in-situ/environmental experiments

Limiting factors in single particle cryo electron tomography

Directory of Open Access Journals (Sweden)

Mikhail Kudryashev

2012-07-01

Full Text Available Modern methods of cryo electron microscopy and tomography allow visualization of protein nanomachines in their native state at the nanometer scale. Image processing methods including sub-volume averaging applied to repeating macromolecular elements within tomograms allow exploring their structures within the native context of the cell, avoiding the need for protein isolation and purification. Today, many different data acquisition protocols and software solutions are available to researchers to determine average structures of macromolecular complexes and potentially to classify structural intermediates. Here, we list the density maps reported in the literature, and analyze each structure for the chosen instrumental settings, sample conditions, main processing steps, and obtained resolution. We present conclusions that identify factors currently limiting the resolution gained by this approach.

A national facility for biological cryo-electron microscopy

International Nuclear Information System (INIS)

Saibil, Helen R.; Grünewald, Kay; Stuart, David I.

2015-01-01

This review provides a brief update on the use of cryo-electron microscopy for integrated structural biology, along with an overview of the plans for the UK national facility for electron microscopy being built at the Diamond synchrotron. Three-dimensional electron microscopy is an enormously powerful tool for structural biologists. It is now able to provide an understanding of the molecular machinery of cells, disease processes and the actions of pathogenic organisms from atomic detail through to the cellular context. However, cutting-edge research in this field requires very substantial resources for equipment, infrastructure and expertise. Here, a brief overview is provided of the plans for a UK national three-dimensional electron-microscopy facility for integrated structural biology to enable internationally leading research on the machinery of life. State-of-the-art equipment operated with expert support will be provided, optimized for both atomic-level single-particle analysis of purified macromolecules and complexes and for tomography of cell sections. The access to and organization of the facility will be modelled on the highly successful macromolecular crystallography (MX) synchrotron beamlines, and will be embedded at the Diamond Light Source, facilitating the development of user-friendly workflows providing near-real-time experimental feedback

3D structure of eukaryotic flagella in a quiescent state revealed by cryo-electron tomography

Science.gov (United States)

Nicastro, Daniela; McIntosh, J. Richard; Baumeister, Wolfgang

2005-01-01

We have used cryo-electron tomography to investigate the 3D structure and macromolecular organization of intact, frozen-hydrated sea urchin sperm flagella in a quiescent state. The tomographic reconstructions provide information at a resolution better than 6 nm about the in situ arrangements of macromolecules that are key for flagellar motility. We have visualized the heptameric rings of the motor domains in the outer dynein arm complex and determined that they lie parallel to the plane that contains the axes of neighboring flagellar microtubules. Both the material associated with the central pair of microtubules and the radial spokes display a plane of symmetry that helps to explain the planar beat pattern of these flagella. Cryo-electron tomography has proven to be a powerful technique for helping us understand the relationships between flagellar structure and function and the design of macromolecular machines in situ. PMID:16246999

Alignment of cryo-EM movies of individual particles by optimization of image translations.

Science.gov (United States)

Rubinstein, John L; Brubaker, Marcus A

2015-11-01

Direct detector device (DDD) cameras have revolutionized single particle electron cryomicroscopy (cryo-EM). In addition to an improved camera detective quantum efficiency, acquisition of DDD movies allows for correction of movement of the specimen, due to both instabilities in the microscope specimen stage and electron beam-induced movement. Unlike specimen stage drift, beam-induced movement is not always homogeneous within an image. Local correlation in the trajectories of nearby particles suggests that beam-induced motion is due to deformation of the ice layer. Algorithms have already been described that can correct movement for large regions of frames and for >1 MDa protein particles. Another algorithm allows individual images to be aligned without frame averaging or linear trajectories. The algorithm maximizes the overall correlation of the shifted frames with the sum of the shifted frames. The optimum in this single objective function is found efficiently by making use of analytically calculated derivatives of the function. To smooth estimates of particle trajectories, rapid changes in particle positions between frames are penalized in the objective function and weighted averaging of nearby trajectories ensures local correlation in trajectories. This individual particle motion correction, in combination with weighting of Fourier components to account for increasing radiation damage in later frames, can be used to improve 3-D maps from single particle cryo-EM. Copyright © 2015 Elsevier Inc. All rights reserved.

Restoring defect structures in 3C-SiC/Si (001) from spherical aberration-corrected high-resolution transmission electron microscope images by means of deconvolution processing.

Science.gov (United States)

Wen, C; Wan, W; Li, F H; Tang, D

2015-04-01

The [110] cross-sectional samples of 3C-SiC/Si (001) were observed with a spherical aberration-corrected 300 kV high-resolution transmission electron microscope. Two images taken not close to the Scherzer focus condition and not representing the projected structures intuitively were utilized for performing the deconvolution. The principle and procedure of image deconvolution and atomic sort recognition are summarized. The defect structure restoration together with the recognition of Si and C atoms from the experimental images has been illustrated. The structure maps of an intrinsic stacking fault in the area of SiC, and of Lomer and 60° shuffle dislocations at the interface have been obtained at atomic level. Copyright © 2015 Elsevier Ltd. All rights reserved.

Imaging of Norway spruce early somatic embryos with the ESEM, Cryo-SEM and laser scanning microscope.

Science.gov (United States)

Neděla, Vilém; Hřib, Jiří; Havel, Ladislav; Hudec, Jiří; Runštuk, Jiří

2016-05-01

This article describes the surface structure of Norway spruce early somatic embryos (ESEs) as a typical culture with asynchronous development. The microstructure of extracellular matrix covering ESEs were observed using the environmental scanning electron microscope as a primary tool and using the scanning electron microscope with cryo attachment and laser electron microscope as a complementary tool allowing our results to be proven independently. The fresh samples were observed in conditions of the air environment of the environmental scanning electron microscope (ESEM) with the pressure from 550Pa to 690Pa and the low temperature of the sample from -18°C to -22°C. The samples were studied using two different types of detector to allow studying either the thin surface structure or material composition. The scanning electron microscope with cryo attachment was used for imaging frozen extracellular matrix microstructure with higher resolution. The combination of both electron microscopy methods was suitable for observation of "native" plant samples, allowing correct evaluation of our results, free of error and artifacts. Copyright © 2016 Elsevier Ltd. All rights reserved.

From Nanoparticles to Process An Aberration Corrected TEM Study of Fischer Tropsch Catalysts at Various Steps of the Process

International Nuclear Information System (INIS)

Braidy, N.; Blanchard, J.; Abatzoglou, N.; Andrei, C.

2011-01-01

χThe nanostructure of Fischer-Tropsch (FT) Fe carbides are investigated using aberration-corrected high-resolution transmission electron microscopy (TEM). The plasma-generated Fe carbides are analyzed just after synthesis, following reduction via a H2 treatment step and once used as FT catalyst and deactivated. The as-produced nanoparticles (NPs) are seen to be abundantly covered with graphitic and amorphous carbon. Using the extended information limit from the spherical aberration-corrected TEM, the NPs could be indexed as a mixture of NPs in the θ-Fe 3 C and χ-Fe 5 C 2 phases. The reduction treatment exposed the NPs by removing most of the carbonaceous speSubscript textcies while retaining the χ-Fe 5 C 2 . Fe-carbides NPs submitted to conditions typical to FT synthesis develop a Fe3O4 shell which eventually consumes the NPs up to a point where 3-4 nm residual carbide is left at the center of the particle. Subscript textVarious mechanisms explaining the formation of such a microstructure are discussed. (author)

Aberration corrected STEM of defects in epitaxial n=4 Ruddlesden-Popper phase Can+1MnnO3n+1

International Nuclear Information System (INIS)

Wang, P; Bleloch, A L; Goodhew, P J; Yan, L; Niu, H J; Rosseinsky, M J; Chalker, P R

2008-01-01

Defects in Ruddlesden-Popper phase CaO·[(CaMnO 3 )] 4 epitaxial films grown on SrTiO 3 (001) by pulsed laser deposition have been investigated using high angle annular dark field imaging in an aberration-corrected STEM. The stacking faults perpendicular and parallel to the substrate formed during the growth are discussed in detail. The desired n = 4 RP phase is imaged and chemically analyzed at the atomic scale using electron energy loss spectroscopy.

Investigating the chemical and morphological evolution of GaAs capped InAs/InP quantum dots emitting at 1.5μm using aberration-corrected scanning transmission electron microscopy

DEFF Research Database (Denmark)

Kadkhodazadeh, Shima; Semenova, Elizaveta; Yvind, Kresten

2011-01-01

The emission wavelength of InAs quantum dots grown on InP has been shown to shift to the technologically desirable 1.5μm with the deposition of 1–2 monolayers of GaAs on top of the quantum dots. Here, we use aberration-corrected scanning transmission electron microscopy to investigate morphological...... and compositional changes occurring to the quantum dots as a result of the deposition of 1.7 monolayers of GaAs on top of them, prior to complete overgrowth with InP. The results are compared with theoretical models describing the overgrowth process....

CryoEM at IUCrJ: a new era

Directory of Open Access Journals (Sweden)

Sriram Subramaniam

2016-01-01

Full Text Available In this overview, we briefly outline recent advances in electron cryomicroscopy (cryoEM and explain why the journal IUCrJ, published by the International Union of Crystallography, could provide a natural home for publications covering many present and future developments in the cryoEM field.

Joint denoising, demosaicing, and chromatic aberration correction for UHD video

Science.gov (United States)

Jovanov, Ljubomir; Philips, Wilfried; Damstra, Klaas Jan; Ellenbroek, Frank

2017-09-01

High-resolution video capture is crucial for numerous applications such as surveillance, security, industrial inspection, medical imaging and digital entertainment. In the last two decades, we are witnessing a dramatic increase of the spatial resolution and the maximal frame rate of video capturing devices. In order to achieve further resolution increase, numerous challenges will be facing us. Due to the reduced size of the pixel, the amount of light also reduces, leading to the increased noise level. Moreover, the reduced pixel size makes the lens imprecisions more pronounced, which especially applies to chromatic aberrations. Even in the case when high quality lenses are used some chromatic aberration artefacts will remain. Next, noise level additionally increases due to the higher frame rates. To reduce the complexity and the price of the camera, one sensor captures all three colors, by relying on Color Filter Arrays. In order to obtain full resolution color image, missing color components have to be interpolated, i.e. demosaicked, which is more challenging than in the case of lower resolution, due to the increased noise and aberrations. In this paper, we propose a new method, which jointly performs chromatic aberration correction, denoising and demosaicking. By jointly performing the reduction of all artefacts, we are reducing the overall complexity of the system and the introduction of new artefacts. In order to reduce possible flicker we also perform temporal video enhancement. We evaluate the proposed method on a number of publicly available UHD sequences and on sequences recorded in our studio.

The 2017 Nobel Prize in Chemistry: cryo-EM comes of age.

Science.gov (United States)

Shen, Peter S

2018-03-01

The 2017 Nobel Prize in Chemistry was awarded to Jacques Dubochet, Joachim Frank, and Richard Henderson for "developing cryo-electron microscopy (cryo-EM) for the high-resolution structure determination of biomolecules in solution." This feature article summarizes some of the major achievements leading to the development of cryo-EM and recent technological breakthroughs that have transformed the method into a mainstream tool for structure determination.

Electron Optics for Biologists: Physical Origins of Spherical Aberrations

Science.gov (United States)

Geissler, Peter; Zadunaisky, Jose

1974-01-01

Reports on the physical origins of spherical aberrations in axially symmetric electrostatic lenses to convey the essentials of electon optics to those who must think critically about the resolution of the electron microscope. (GS)

Characterization of intact subcellular bodies in whole bacteria by cryo-electron tomography and spectroscopic imaging.

Science.gov (United States)

Comolli, L R; Kundmann, M; Downing, K H

2006-07-01

We illustrate the combined use of cryo-electron tomography and spectroscopic difference imaging in the study of subcellular structure and subcellular bodies in whole bacteria. We limited our goal and focus to bodies with a distinct elemental composition that was in a sufficiently high concentration to provide the necessary signal-to-noise level at the relatively large sample thicknesses of the intact cell. This combination proved very powerful, as demonstrated by the identification of a phosphorus-rich body in Caulobacter crescentus. We also confirmed the presence of a body rich in carbon, demonstrated that these two types of bodies are readily recognized and distinguished from each other, and provided, for the first time to our knowledge, structural information about them in their intact state. In addition, we also showed the presence of a similar type of phosphorus-rich body in Deinococcus grandis, a member of a completely unrelated bacteria genus. Cryo-electron microscopy and tomography allowed the study of the biogenesis and morphology of these bodies at resolutions better than 10 nm, whereas spectroscopic difference imaging provided a direct identification of their chemical composition.

Recent advances in cryo-TEM imaging of soft lipid nanoparticles

DEFF Research Database (Denmark)

Helvig, Shen Yu; Mat Azmi, Intan Diana Binti; Moghimi, Seyed Moien

2015-01-01

Cryo-transmission electron microscopy (Cryo-TEM), and its technological variations thereof, have become a powerful tool for detailed morphological characterization and 3D tomography of soft lipid and polymeric nanoparticles as well as biological materials such as viruses and DNA without chemical...... fixation. Here, we review and discuss recent advances in Cryo-TEM analysis of lipid-based drug nanocarriers with particular emphasis on morphological and internal nanostructure characterization of lyotropic liquid crystalline nanoparticles such as cubosomes and hexosomes....

The influence of C{sub s}/C{sub c} correction in analytical imaging and spectroscopy in scanning and transmission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Zaluzec, Nestor J., E-mail: zaluzec@microscopy.com

2015-04-15

Aberration correction in scanning/transmission electron microscopy (S/TEM) owes much to the efforts of a small dedicated group of innovators. Leading that frontier has been Prof. Harald Rose. To date his leadership and dynamic personality has spearheaded our ability to leave behind many of the limitations imposed by spherical aberration (C{sub s}) in high resolution phase contrast imaging. Following shortly behind, has been the development of chromatic aberration correction (C{sub c}) which augments those accomplishments. In this paper we will review and summarize how the combination of C{sub s}/C{sub c} technology enhances our ability to conduct hyperspectral imaging and spectroscopy in today's and future computationally mediated experiments in both thin as well as realistic specimens in vacuo and during in-situ/environmental experiments.

Zernike Phase Contrast Electron Cryo-Tomography Applied to Marine Cyanobacteria Infected with Cyanophages

Science.gov (United States)

Dai, Wei; Fu, Caroline; Khant, Htet A.; Ludtke, Steven J.; Schmid, Michael F.; Chiu, Wah

2015-01-01

Advances in electron cryo-tomography have provided a new opportunity to visualize the internal 3D structures of a bacterium. An electron microscope equipped with Zernike phase contrast optics produces images with dramatically increased contrast compared to images obtained by conventional electron microscopy. Here we describe a protocol to apply Zernike phase plate technology for acquiring electron tomographic tilt series of cyanophage-infected cyanobacterial cells embedded in ice, without staining or chemical fixation. We detail the procedures for aligning and assessing phase plates for data collection, and methods to obtain 3D structures of cyanophage assembly intermediates in the host, by subtomogram alignment, classification and averaging. Acquiring three to four tomographic tilt series takes approximately 12 h on a JEM2200FS electron microscope. We expect this time requirement to decrease substantially as the technique matures. Time required for annotation and subtomogram averaging varies widely depending on the project goals and data volume. PMID:25321408

Camera processing with chromatic aberration.

Science.gov (United States)

Korneliussen, Jan Tore; Hirakawa, Keigo

2014-10-01

Since the refractive index of materials commonly used for lens depends on the wavelengths of light, practical camera optics fail to converge light to a single point on an image plane. Known as chromatic aberration, this phenomenon distorts image details by introducing magnification error, defocus blur, and color fringes. Though achromatic and apochromatic lens designs reduce chromatic aberration to a degree, they are complex and expensive and they do not offer a perfect correction. In this paper, we propose a new postcapture processing scheme designed to overcome these problems computationally. Specifically, the proposed solution is comprised of chromatic aberration-tolerant demosaicking algorithm and post-demosaicking chromatic aberration correction. Experiments with simulated and real sensor data verify that the chromatic aberration is effectively corrected.

CryoSat/SIRAL Cal1 Calibration Orbits

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; Bouffard, Jerome; Parrinello, Tommaso

2017-04-01

The main payload of CryoSat is a Ku band pulsewidth limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for SAR processing. This allows to reach an along track resolution that is significantly improved with respect to traditional pulse-width limited altimeters. Due to the fact that SIRAL is a phase coherent pulse-width limited radar altimeter, a proper calibration approach has been developed. In fact, not only corrections for transfer function, gain and instrument path delay have to be computed (as in previous altimeters), but also corrections for phase (SAR/SARIn) and phase difference between the two receiving chains (SARIN only). Recalling that the CryoSat's orbit has a high inclination of 92° and it is non-sun-synchronous, the temperature of the SIRAL changes continuously along the orbit with a period of about 480 days and it is also function of the ascending/descending passes. By analysis of the CAL1 calibration corrections, it has been verified that the internal path delay and the instrument gain variation measured on the SIRAL are affected by the thermal status of the instrument and as a consequence they are expected to vary along the orbit. In order to gain knowledge on the calibration corrections (i.e. the instrument behavior) as function of latitude and temperature, it has been planned to command a few number of orbits where only CAL1 calibration acquisitions are continuously performed. The analysis of the CAL1 calibration corrections produced along the Calibration orbits can be also useful to verify whether the current calibration plan is able to provide sufficiently accurate corrections for the instrument acquisitions at any latitude. In 2016, the CryoSat/SIRAL Cal1 Calibration Orbits have been commanded two times, a first time the 20th of July 2016 and a second time the 24th of November 2016, and they

Aberration compensation of an ultrasound imaging instrument with a reduced number of channels.

Science.gov (United States)

Jiang, Wei; Astheimer, Jeffrey P; Waag, Robert C

2012-10-01

Focusing and imaging qualities of an ultrasound imaging system that uses aberration correction were experimentally investigated as functions of the number of parallel channels. Front-end electronics that consolidate signals from multiple physical elements can be used to lower hardware and computational costs by reducing the number of parallel channels. However, the signals from sparse arrays of synthetic elements yield poorer aberration estimates. In this study, aberration estimates derived from synthetic arrays of varying element sizes are evaluated by comparing compensated receive focuses, compensated transmit focuses, and compensated b-scan images of a point target and a cyst phantom. An array of 80 x 80 physical elements with a pitch of 0.6 x 0.6 mm was used for all of the experiments and the aberration was produced by a phantom selected to mimic propagation through abdominal wall. The results show that aberration correction derived from synthetic arrays with pitches that have a diagonal length smaller than 70% of the correlation length of the aberration yield focuses and images of approximately the same quality. This connection between correlation length of the aberration and synthetic element size provides a guideline for determining the number of parallel channels that are required when designing imaging systems that employ aberration correction.

Aberration-corrected scanning transmission electron microscopy of semiconductors

International Nuclear Information System (INIS)

Krivanek, O L; Dellby, N; Murfitt, M F

2011-01-01

The scanning transmission electron microscope (STEM) has been able to image individual heavy atoms in a light matrix for some time. It is now able to do much more: it can resolve individual atoms as light as boron in monolayer materials; image atomic columns as light as hydrogen, identify the chemical type of individual isolated atoms from the intensity of their annular dark field (ADF) image and by electron energy loss spectroscopy (EELS); and map elemental composition at atomic resolution by EELS and energy-dispersive X-ray spectroscopy (EDXS). It can even map electronic states, also by EELS, at atomic resolution. The instrumentation developments that have made this level of performance possible are reviewed, and examples of applications to semiconductors and oxides are shown.

Single-particle cryo-electron microscopy of Rift Valley fever virus

International Nuclear Information System (INIS)

Sherman, Michael B.; Freiberg, Alexander N.; Holbrook, Michael R.; Watowich, Stanley J.

2009-01-01

Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human and veterinary pathogen causing acute hepatitis in ruminants and has the potential to cause hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on the virus surface are highly symmetric and arranged on a T = 12 icosahedral lattice. Our RVFV MP-12 structure allowed clear identification of inter-capsomer contacts and definition of possible glycoprotein arrangements within capsomers. This structure provides a detailed model for phleboviruses, opens new avenues for high-resolution structural studies of the bunyavirus family, and aids the design of antiviral diagnostics and effective subunit vaccines.

Single-particle cryo-electron microscopy of Rift Valley fever virus.

Science.gov (United States)

Sherman, Michael B; Freiberg, Alexander N; Holbrook, Michael R; Watowich, Stanley J

2009-04-25

Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human and veterinary pathogen causing acute hepatitis in ruminants and has the potential to cause hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on the virus surface are highly symmetric and arranged on a T=12 icosahedral lattice. Our RVFV MP-12 structure allowed clear identification of inter-capsomer contacts and definition of possible glycoprotein arrangements within capsomers. This structure provides a detailed model for phleboviruses, opens new avenues for high-resolution structural studies of the bunyavirus family, and aids the design of antiviral diagnostics and effective subunit vaccines.

Optical traps with geometric aberrations

International Nuclear Information System (INIS)

Roichman, Yael; Waldron, Alex; Gardel, Emily; Grier, David G.

2006-01-01

We assess the influence of geometric aberrations on the in-plane performance of optical traps by studying the dynamics of trapped colloidal spheres in deliberately distorted holographic optical tweezers. The lateral stiffness of the traps turns out to be insensitive to moderate amounts of coma, astigmatism, and spherical aberration. Moreover holographic aberration correction enables us to compensate inherent shortcomings in the optical train, thereby adaptively improving its performance. We also demonstrate the effects of geometric aberrations on the intensity profiles of optical vortices, whose readily measured deformations suggest a method for rapidly estimating and correcting geometric aberrations in holographic trapping systems

In-situ Study of Dynamic Phenomena at Metal Nanosolder Interfaces Using Aberration Corrected Scanning Transmission Electron Microcopy.

Energy Technology Data Exchange (ETDEWEB)

Lu, Ping

2014-10-01

Controlling metallic nanoparticle (NP) interactions plays a vital role in the development of new joining techniques (nanosolder) that bond at lower processing temperatures but remain viable at higher temperatures. The pr imary objective of this project is t o develop a fundamental understanding of the actual reaction processes, associated atomic mechanisms, and the resulting microstructure that occur during thermally - driven bond formation concerning metal - metal nano - scale (%3C50nm) interfaces. In this LDRD pr oject, we have studied metallic NPs interaction at the elevated temperatures by combining in - situ transmission electron microscopy (TEM ) using an aberration - corrected scanning transmission electron microscope (AC - STEM) and atomic - scale modeling such as m olecular dynamic (MD) simulations. Various metallic NPs such as Ag, Cu and Au are synthesized by chemical routines. Numerous in - situ e xperiments were carried out with focus of the research on study of Ag - Cu system. For the first time, using in - situ STEM he ating experiments , we directly observed t he formation of a 3 - dimensional (3 - D) epitaxial Cu - Ag core - shell nanoparticle during the thermal interaction of Cu and Ag NPs at elevated temperatures (150 - 300 o C). The reaction takes place at temperatures as low as 150 o C and was only observed when care was taken to circumvent the effects of electron beam irradiation during STEM imaging. Atomic - scale modeling verified that the Cu - Ag core - shell structure is energetically favored, and indicated that this phenomenon is a nano - scale effect related to the large surface - to - volume ratio of the NPs. The observation potentially can be used for developing new nanosolder technology that uses Ag shell as the "glue" that stic ks the particles of Cu together. The LDRD has led to several journal publications and numerous conference presentations, and a TA. In addition, we have developed new TEM characterization techniques and phase

Exploring an optimal wavelet-based filter for cryo-ET imaging.

Science.gov (United States)

Huang, Xinrui; Li, Sha; Gao, Song

2018-02-07

Cryo-electron tomography (cryo-ET) is one of the most advanced technologies for the in situ visualization of molecular machines by producing three-dimensional (3D) biological structures. However, cryo-ET imaging has two serious disadvantages-low dose and low image contrast-which result in high-resolution information being obscured by noise and image quality being degraded, and this causes errors in biological interpretation. The purpose of this research is to explore an optimal wavelet denoising technique to reduce noise in cryo-ET images. We perform tests using simulation data and design a filter using the optimum selected wavelet parameters (three-level decomposition, level-1 zeroed out, subband-dependent threshold, a soft-thresholding and spline-based discrete dyadic wavelet transform (DDWT)), which we call a modified wavelet shrinkage filter; this filter is suitable for noisy cryo-ET data. When testing using real cryo-ET experiment data, higher quality images and more accurate measures of a biological structure can be obtained with the modified wavelet shrinkage filter processing compared with conventional processing. Because the proposed method provides an inherent advantage when dealing with cryo-ET images, it can therefore extend the current state-of-the-art technology in assisting all aspects of cryo-ET studies: visualization, reconstruction, structural analysis, and interpretation.

Wavefront correction for static and dynamic aberrations to within 1 second of the system shot in the NIF Beamlet demonstration facility

International Nuclear Information System (INIS)

Hartley, R.; Kartz, M.; Behrendt, W.

1996-10-01

The laser wavefront of the NIF Beamlet demonstration system is corrected for static aberrations with a wavefront control system. The system operates closed loop with a probe beam prior to a shot and has a loop bandwidth of about 3 Hz. However, until recently the wavefront control system was disabled several minutes prior to the shot to allow time to manually reconfigure its attenuators and probe beam insertion mechanism to shot mode. Thermally-induced dynamic variations in gas density in the Beamlet main beam line produce significant wavefront error. After about 5-8 seconds, the wavefront error has increased to a new, higher level due to turbulence- induced aberrations no longer being corrected- This implies that there is a turbulence-induced aberration noise bandwidth of less than one Hertz, and that the wavefront controller could correct for the majority of turbulence-induced aberration (about one- third wave) by automating its reconfiguration to occur within one second of the shot, This modification was recently implemented on Beamlet; we call this modification the t 0 -1 system

Aberration Correction in the Brewer Spectrophotometer

International Nuclear Information System (INIS)

Johnston, J.E.; Kerr, J.B.; McElroy, C.T.; Wardle, D.I.

2000-01-01

The optical design of the Brewer Spectrophotometer has been optimised for measurements in the 300-320 nm wavelength range. An aberration resolution limit that is much less than the 0.6 nm FWHM (full width at half maximum) is achieved by using an Ebert-Fastie spectrometer design, modified by the inclusion tilted lens that optimises performance at 310 nm. The small contribution of the remaining aberration to the measured instrument function is critical to radiometric measurement quality. Ramifications of this design to the development of instrumentation with enhanced scanning abilities are discussed. (author)

Lowered threshold energy for femtosecond laser induced optical breakdown in a water based eye model by aberration correction with adaptive optics.

Science.gov (United States)

Hansen, Anja; Géneaux, Romain; Günther, Axel; Krüger, Alexander; Ripken, Tammo

2013-06-01

In femtosecond laser ophthalmic surgery tissue dissection is achieved by photodisruption based on laser induced optical breakdown. In order to minimize collateral damage to the eye laser surgery systems should be optimized towards the lowest possible energy threshold for photodisruption. However, optical aberrations of the eye and the laser system distort the irradiance distribution from an ideal profile which causes a rise in breakdown threshold energy even if great care is taken to minimize the aberrations of the system during design and alignment. In this study we used a water chamber with an achromatic focusing lens and a scattering sample as eye model and determined breakdown threshold in single pulse plasma transmission loss measurements. Due to aberrations, the precise lower limit for breakdown threshold irradiance in water is still unknown. Here we show that the threshold energy can be substantially reduced when using adaptive optics to improve the irradiance distribution by spatial beam shaping. We found that for initial aberrations with a root-mean-square wave front error of only one third of the wavelength the threshold energy can still be reduced by a factor of three if the aberrations are corrected to the diffraction limit by adaptive optics. The transmitted pulse energy is reduced by 17% at twice the threshold. Furthermore, the gas bubble motions after breakdown for pulse trains at 5 kilohertz repetition rate show a more transverse direction in the corrected case compared to the more spherical distribution without correction. Our results demonstrate how both applied and transmitted pulse energy could be reduced during ophthalmic surgery when correcting for aberrations. As a consequence, the risk of retinal damage by transmitted energy and the extent of collateral damage to the focal volume could be minimized accordingly when using adaptive optics in fs-laser surgery.

CryoVEx 2011-12 Airborne Campaigns for CryoSat Validation

DEFF Research Database (Denmark)

Skourup, Henriette; Hvidegaard, Sine Munk; Forsberg, René

2013-01-01

After the successful launch of CryoSat-2 in April 2010, the first direct validation campaign of the satellite was carried out in the April-May 2011. Part of this was repeated in Spring 2012. DTU Space has been involved in ESA’s CryoSat Validation Experiment (CryoVEx) with airborne activities since...... 2003. To validate the performance of the CryoSat-2 radar altimeter (SIRAL), the aircraft is equipped with an airborne version of the SIRAL altimeter (ASIRAS) together with a laser scanner. Of particular interest is to study the penetration depth of SIRAL into both land- and sea ice. This can be done...... of Alert and sea ice around Svalbard in the Fram Strait. Selected tracks were planned to match CryoSat-2 passes and a few of them were flown in formation flight with the AlfredWegener Institute (AWI) Polar- 5 carrying an EM induction sounder. The paper presents an overview of the 2011-12 airborne campaigns...

Automatic low-order aberration correction based on geometrical optics for slab lasers.

Science.gov (United States)

Yu, Xin; Dong, Lizhi; Lai, Boheng; Yang, Ping; Liu, Yong; Kong, Qingfeng; Yang, Kangjian; Tang, Guomao; Xu, Bing

2017-02-20

In this paper, we present a method based on geometry optics to simultaneously correct low-order aberrations and reshape the beams of slab lasers. A coaxial optical system with three lenses is adapted. The positions of the three lenses are directly calculated based on the beam parameters detected by wavefront sensors. The initial sizes of the input beams are 1.8  mm×11  mm, and peak-to-valley (PV) values of the wavefront range up to several tens of microns. After automatic correction, the dimensions may reach nearly 22  mm×22  mm as expected, and PV values of the wavefront are less than 2 μm. The effectiveness and precision of this method are verified with experiments.

Spatial Localization of the Ebola Virus Glycoprotein Mucin-Like Domain Determined by Cryo-Electron Tomography

OpenAIRE

Tran, Erin E. H.; Simmons, James A.; Bartesaghi, Alberto; Shoemaker, Charles J.; Nelson, Elizabeth; White, Judith M.; Subramaniam, Sriram

2014-01-01

The Ebola virus glycoprotein mucin-like domain (MLD) is implicated in Ebola virus cell entry and immune evasion. Using cryo-electron tomography of Ebola virus-like particles, we determined a three-dimensional structure for the full-length glycoprotein in a near-native state and compared it to that of a glycoprotein lacking the MLD. Our results, which show that the MLD is located at the apex and the sides of each glycoprotein monomer, provide a structural template for analysis of MLD function.

Coastal Sea Level from CryoSat-2 SARIn Altimetry in Norway

DEFF Research Database (Denmark)

Idžanović, Martina; Ophaug, Vegard; Andersen, Ole Baltazar

2017-01-01

Conventional (pulse-limited) altimeters determine the sea surface height with an accuracy of a few centimeters over the open ocean. Sea surface heights and tide-gauge sea level serve as each other’s buddy check. However, in coastal areas, altimetry suffers from numerous effects, which degrade its...... conventional altimeters. In this study, we explore the potential of CryoSat-2 to provide valid observations in the Norwegian coastal zone. We do this by comparing time series of CryoSat-2 sea level anomalies with time series of in situ sea level at 22 tide gauges, where the CryoSat-2 sea level anomalies...... are averaged in a 45-km area around each tide gauge. For all tide gauges, CryoSat-2 shows standard deviations of differences and correlations of 16 cm and 61%, respectively. We further identify the ocean tide and inverted barometer geophysical corrections as the most crucial, and note that a large amount...

Ultra-precise measurement of optical aberrations for sub-Aangstroem transmission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Barthel, J.

2008-06-15

Quantitative investigations of material structures on an atomic scale by means of highresolution transmission electron microscopy (HRTEM) impose not only extreme demands on the mechanic and electromagnetic stability of the applied instruments but require also their precise electron-optical adjustment. Today a physical resolution well below one Aangstroem can be achieved with commercially available microscopes on a daily basis. However, the achieved resolution can often not be reliably exploited for the interpretation of the resulting microscopical data due to the presence of so-called higher-order lens aberrations. At the starting time of this work, a sufficiently accurate procedure to measure higher-order aberrations was urgently missing. Since aberration measurement is a mandatory prerequisite for any technique of aberration control enabling quantitative high-resolution microscopy, the goal of this work is to develop such a measurement procedure for the Sub-Aangstroem regime. The measurement procedures developed in the course of this work are based on the numerical evaluation of a series of images taken from an amorphous object under electron-beam illumination with varying tilt. New techniques have been developed for the evaluation of single images as well as for the optimised evaluation of the whole series. These procedures allow microscope users to perform quantitative HRTEM even at a resolution of 0.5 Aangstroem. The precision reached with the newly developed measurement procedures is unprecedented and surpasses existing solutions by at least one order of magnitude in any respect. All the concepts and procedures for aberration measurement developed in this work have been implemented in a software package which satisfies professional demands with respect to robustness, precision, speed and user-friendliness. The new automatic aberrationmeasurement procedures are suitable to establish HRTEM as a quantitative technique for material science investigations in the

Ultra-precise measurement of optical aberrations for sub-Aangstroem transmission electron microscopy

International Nuclear Information System (INIS)

Barthel, J.

2008-06-01

Quantitative investigations of material structures on an atomic scale by means of highresolution transmission electron microscopy (HRTEM) impose not only extreme demands on the mechanic and electromagnetic stability of the applied instruments but require also their precise electron-optical adjustment. Today a physical resolution well below one Aangstroem can be achieved with commercially available microscopes on a daily basis. However, the achieved resolution can often not be reliably exploited for the interpretation of the resulting microscopical data due to the presence of so-called higher-order lens aberrations. At the starting time of this work, a sufficiently accurate procedure to measure higher-order aberrations was urgently missing. Since aberration measurement is a mandatory prerequisite for any technique of aberration control enabling quantitative high-resolution microscopy, the goal of this work is to develop such a measurement procedure for the Sub-Aangstroem regime. The measurement procedures developed in the course of this work are based on the numerical evaluation of a series of images taken from an amorphous object under electron-beam illumination with varying tilt. New techniques have been developed for the evaluation of single images as well as for the optimised evaluation of the whole series. These procedures allow microscope users to perform quantitative HRTEM even at a resolution of 0.5 Aangstroem. The precision reached with the newly developed measurement procedures is unprecedented and surpasses existing solutions by at least one order of magnitude in any respect. All the concepts and procedures for aberration measurement developed in this work have been implemented in a software package which satisfies professional demands with respect to robustness, precision, speed and user-friendliness. The new automatic aberrationmeasurement procedures are suitable to establish HRTEM as a quantitative technique for material science investigations in the

On the optical stability of high-resolution transmission electron microscopes

International Nuclear Information System (INIS)

Barthel, J.; Thust, A.

2013-01-01

In the recent two decades the technique of high-resolution transmission electron microscopy experienced an unprecedented progress through the introduction of hardware aberration correctors and by the improvement of the achievable resolution to the sub-Ångström level. The important aspect that aberration correction at a given resolution requires also a well defined amount of optical stability has received little attention so far. Therefore we investigate the qualification of a variety of high-resolution electron microscopes to maintain an aberration corrected optical state in terms of an optical lifetime. We develop a comprehensive statistical framework for the estimation of the optical lifetime and find remarkably low values between tens of seconds and a couple of minutes. Probability curves are introduced, which inform the operator about the chance to work still in the fully aberration corrected state. - Highlights: • We investigate the temporal stability of optical aberrations in HRTEM. • We develop a statistical framework for the estimation of optical lifetimes. • We introduce plots showing the success probability for aberration-free work. • Optical lifetimes in sub-Ångström electron microscopy are surprisingly low. • The success of aberration correction depends strongly on the optical stability

Unsupervised Cryo-EM Data Clustering through Adaptively Constrained K-Means Algorithm.

Science.gov (United States)

Xu, Yaofang; Wu, Jiayi; Yin, Chang-Cheng; Mao, Youdong

2016-01-01

In single-particle cryo-electron microscopy (cryo-EM), K-means clustering algorithm is widely used in unsupervised 2D classification of projection images of biological macromolecules. 3D ab initio reconstruction requires accurate unsupervised classification in order to separate molecular projections of distinct orientations. Due to background noise in single-particle images and uncertainty of molecular orientations, traditional K-means clustering algorithm may classify images into wrong classes and produce classes with a large variation in membership. Overcoming these limitations requires further development on clustering algorithms for cryo-EM data analysis. We propose a novel unsupervised data clustering method building upon the traditional K-means algorithm. By introducing an adaptive constraint term in the objective function, our algorithm not only avoids a large variation in class sizes but also produces more accurate data clustering. Applications of this approach to both simulated and experimental cryo-EM data demonstrate that our algorithm is a significantly improved alterative to the traditional K-means algorithm in single-particle cryo-EM analysis.

Investigations in space-related molecular biology. [cryo-electron microscopic and diffraction studies on terrestrial and extraterrestrial specimens

Science.gov (United States)

Fernandez-Moran, H.; Pritzker, A. N.

1974-01-01

Improved instrumentation and preparation techniques for high resolution, high voltage cryo-electron microscopic and diffraction studies on terrestrial and extraterrestrial specimens are reported. Computer correlated ultrastructural and biochemical work on hydrated and dried cell membranes and related biological systems provided information on membrane organization, ice crystal formation and ordered water, RNA virus linked to cancer, lunar rock samples, and organometallic superconducting compounds. Apollo 11, 12, 14, and 15 specimens were analyzed

(CryoTransmission Electron Microscopy of Phospholipid Model Membranes Interacting with Amphiphilic and Polyphilic Molecules

Directory of Open Access Journals (Sweden)

Annette Meister

2017-10-01

Full Text Available Lipid membranes can incorporate amphiphilic or polyphilic molecules leading to specific functionalities and to adaptable properties of the lipid bilayer host. The insertion of guest molecules into membranes frequently induces changes in the shape of the lipid matrix that can be visualized by transmission electron microscopy (TEM techniques. Here, we review the use of stained and vitrified specimens in (cryoTEM to characterize the morphology of amphiphilic and polyphilic molecules upon insertion into phospholipid model membranes. Special emphasis is placed on the impact of novel synthetic amphiphilic and polyphilic bolalipids and polymers on membrane integrity and shape stability.

A comparative analysis of the sea ice freeboard from CryoSat. CryoVEx and IceBridge

DEFF Research Database (Denmark)

Kildegaard Rose, Stine; Connor, Laurence N.; Farrell, Sinead L.

The CryoSat Validation Experiment (CryoVEx) has been conducted by ESA, to examine the uncertainties in the satellite measurement of e.g. sea ice thickness. In this study, we aim to estimate the sea ice freeboard from CryoSat-2, and compare it with the high-resolution Airborne Laser Scanner (ALS......) measurements collected along CryoSat-2 ground tracks from the CryoVEx 2012 campaign, together with NASA’s Operation IceBridge data. We will use the CryoSat SAR data level 1b (L1b) to discriminate the leads and from this, estimate the sea ice freeboard. Furthermore, we are looking at the CryoSat level 2 (L2...

Reply to L.M. Brown et al. “Brief history of the Cambridge STEM aberration correction project and its progeny” in Ultramicroscopy 157, 88 (2015)

Energy Technology Data Exchange (ETDEWEB)

Urban, K.W., E-mail: k.urban@fz-juelich.de [Peter Grünberg Institute, Forschungszentrum Jülich, Jülich-Aachen Research Alliance (JARA), and Ernst Ruska Centre for Microscopy and Spectroscopy with Electrons, 52425 Jülich (Germany); Rose, H. [Materialwissenschaftliche Elektronenmikroskopie, Universität Ulm, Albert-Einstein-Allee 11, 89081 Ulm (Germany)

2016-02-15

We comment on a Short Communication recently published in Ultramicroscopy in which Brown et al. criticize our description of the time sequence of events in the development of aberration correction systems in electron optics during the 1990s put forward in the introduction to the Ultramicroscopy April 2015 Special Issue. We present an analysis of the published literature furnishing evidence that our description is correct. - Highlights: • We scrutinize assertions made on the evolution of Cambridge Cs corrector project. • References [22-24] do not demonstrate improvement of resolution by Cs correction. • According to literature such improvement is only shown in reference [10] in 2001. • Corresponding evidence was published by Heidelberg project already in 1998. • The Heidelberg Cs corrector project antedates the Cambridge project by 3 years.

Reply to L.M. Brown et al. “Brief history of the Cambridge STEM aberration correction project and its progeny” in Ultramicroscopy 157, 88 (2015)

International Nuclear Information System (INIS)

Urban, K.W.; Rose, H.

2016-01-01

We comment on a Short Communication recently published in Ultramicroscopy in which Brown et al. criticize our description of the time sequence of events in the development of aberration correction systems in electron optics during the 1990s put forward in the introduction to the Ultramicroscopy April 2015 Special Issue. We present an analysis of the published literature furnishing evidence that our description is correct. - Highlights: • We scrutinize assertions made on the evolution of Cambridge Cs corrector project. • References [22-24] do not demonstrate improvement of resolution by Cs correction. • According to literature such improvement is only shown in reference [10] in 2001. • Corresponding evidence was published by Heidelberg project already in 1998. • The Heidelberg Cs corrector project antedates the Cambridge project by 3 years.

Spatial localization of the Ebola virus glycoprotein mucin-like domain determined by cryo-electron tomography.

Science.gov (United States)

Tran, Erin E H; Simmons, James A; Bartesaghi, Alberto; Shoemaker, Charles J; Nelson, Elizabeth; White, Judith M; Subramaniam, Sriram

2014-09-01

The Ebola virus glycoprotein mucin-like domain (MLD) is implicated in Ebola virus cell entry and immune evasion. Using cryo-electron tomography of Ebola virus-like particles, we determined a three-dimensional structure for the full-length glycoprotein in a near-native state and compared it to that of a glycoprotein lacking the MLD. Our results, which show that the MLD is located at the apex and the sides of each glycoprotein monomer, provide a structural template for analysis of MLD function. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Statistical estimation of ultrasonic propagation path parameters for aberration correction.

Science.gov (United States)

Waag, Robert C; Astheimer, Jeffrey P

2005-05-01

Parameters in a linear filter model for ultrasonic propagation are found using statistical estimation. The model uses an inhomogeneous-medium Green's function that is decomposed into a homogeneous-transmission term and a path-dependent aberration term. Power and cross-power spectra of random-medium scattering are estimated over the frequency band of the transmit-receive system by using closely situated scattering volumes. The frequency-domain magnitude of the aberration is obtained from a normalization of the power spectrum. The corresponding phase is reconstructed from cross-power spectra of subaperture signals at adjacent receive positions by a recursion. The subapertures constrain the receive sensitivity pattern to eliminate measurement system phase contributions. The recursion uses a Laplacian-based algorithm to obtain phase from phase differences. Pulse-echo waveforms were acquired from a point reflector and a tissue-like scattering phantom through a tissue-mimicking aberration path from neighboring volumes having essentially the same aberration path. Propagation path aberration parameters calculated from the measurements of random scattering through the aberration phantom agree with corresponding parameters calculated for the same aberrator and array position by using echoes from the point reflector. The results indicate the approach describes, in addition to time shifts, waveform amplitude and shape changes produced by propagation through distributed aberration under realistic conditions.

Towards sub-0.5 A electron beams

Energy Technology Data Exchange (ETDEWEB)

Krivanek, O.L.; Nellist, P.D.; Dellby, N.; Murfitt, M.F.; Szilagyi, Z

2003-09-15

In the 4 years since the previous meeting in the SALSA series, aberration correction has progressed from a promising concept to a powerful research tool. We summarize the factors that have enabled 100-120 kV scanning transmission electron microscopes to achieve sub-A resolution, and to increase the current available in an atom-sized probe by a factor of 10 and more. Once C{sub s} is corrected, fifth-order spherical aberration (C{sub 5}) and chromatic aberration (C{sub c}) pose new limits on resolution. We describe a quadrupole/octupole corrector of a new design, which will correct all fifth-order aberrations while introducing less than 0.2 mm of additional C{sub c}. Coupled to an optimized STEM column, the new corrector promises to lead to routine sub-A electron probes at 100 kV, and to sub-0.5 A probes at higher operating voltages.

Morphological evolution of InAs/InP quantum wires through aberration-corrected scanning transmission electron microscopy

Energy Technology Data Exchange (ETDEWEB)

Sales, D L; Molina, S I [Departamento de Ciencia de los Materiales e I. M. y Q. I., Universidad de Cadiz, Campus Rio San Pedro, E-11510 Puerto Real, Cadiz (Spain); Varela, M; Pennycook, S J [Materials Science and Technology Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831 (United States); Galindo, P L [Departamento de Lenguajes y Sistemas Informaticos, Universidad de Cadiz, Campus Rio San Pedro, E-11510 Puerto Real, Cadiz (Spain); Gonzalez, L; Gonzalez, Y [Instituto de Microelectronica de Madrid (CNM, CSIC), Isaac Newton 8, E-28760 Tres Cantos, Madrid (Spain); Fuster, D, E-mail: david.sales@uca.es [UMDO - Unidad Asociada al CSIC-IMM, Instituto de Ciencia de Materiales, Universidad de Valencia, PO Box 22085, 4607 Valencia (Spain)

2010-08-13

Evolution of the size, shape and composition of self-assembled InAs/InP quantum wires through the Stranski-Krastanov transition has been determined by aberration-corrected Z-contrast imaging. High resolution compositional maps of the wires in the initial, intermediate and final formation stages are presented. (001) is the main facet at their very initial stage of formation, which is gradually reduced in favour of {l_brace}114{r_brace} or {l_brace}118{r_brace}, ending with the formation of mature quantum wires with {l_brace}114{r_brace} facets. Significant changes in wire dimensions are measured when varying slightly the amount of InAs deposited. These results are used as input parameters to build three-dimensional models that allow calculation of the strain energy during the quantum wire formation process. The observed morphological evolution is explained in terms of the calculated elastic energy changes at the growth front. Regions of the wetting layer close to the nanostructure perimeters have higher strain energy, causing migration of As atoms towards the quantum wire terraces, where the structure is partially relaxed; the thickness of the wetting layer is reduced in these zones and the island height increases until the (001) facet is removed.

Space charge effects and aberrations on electron pulse compression in a spherical electrostatic capacitor.

Science.gov (United States)

Yu, Lei; Li, Haibo; Wan, Weishi; Wei, Zheng; Grzelakowski, Krzysztof P; Tromp, Rudolf M; Tang, Wen-Xin

2017-12-01

The effects of space charge, aberrations and relativity on temporal compression are investigated for a compact spherical electrostatic capacitor (α-SDA). By employing the three-dimensional (3D) field simulation and the 3D space charge model based on numerical General Particle Tracer and SIMION, we map the compression efficiency for a wide range of initial beam size and single-pulse electron number and determine the optimum conditions of electron pulses for the most effective compression. The results demonstrate that both space charge effects and aberrations prevent the compression of electron pulses into the sub-ps region if the electron number and the beam size are not properly optimized. Our results suggest that α-SDA is an effective compression approach for electron pulses under the optimum conditions. It may serve as a potential key component in designing future time-resolved electron sources for electron diffraction and spectroscopy experiments. Copyright © 2017 Elsevier B.V. All rights reserved.

Higher-order aberrations and best-corrected visual acuity in Native American children with a high prevalence of astigmatism.

Science.gov (United States)

Miller, Joseph M; Harvey, Erin M; Schwiegerling, Jim

2015-08-01

To determine whether higher-order aberrations (HOAs) in children from a highly astigmatic population differ from population norms and whether HOAs are associated with astigmatism and reduced best-corrected visual acuity. Subjects were 218 Tohono O'odham Native American children 5-9 years of age. Noncycloplegic HOA measurements were obtained with a handheld Shack-Hartmann sensor (SHS). Signed (z06s to z14s) and unsigned (z06u to z14u) wavefront aberration Zernike coefficients Z(3,-3) to Z(4,4) were rescaled for a 4 mm diameter pupil and compared to adult population norms. Cycloplegic refraction and best-corrected logMAR letter visual acuity (BCVA) were also measured. Regression analyses assessed the contribution of astigmatism (J0) and HOAs to BCVA. The mean root-mean-square (RMS) HOA of 0.191 ± 0.072 μm was significantly greater than population norms (0.100 ± 0.044 μm). All unsigned HOA coefficients (z06u to z14u) and all signed coefficients except z09s, z10s, and z11s were significantly larger than population norms. Decreased BCVA was associated with astigmatism (J0) and spherical aberration (z12u) but not RMS coma, with the effect of J0 about 4 times as great as z12u. Tohono O'odham children show elevated HOAs compared to population norms. Astigmatism and unsigned spherical aberration are associated with decreased acuity, but the effects of spherical aberration are minimal and not clinically significant. Copyright © 2015 American Association for Pediatric Ophthalmology and Strabismus. Published by Elsevier Inc. All rights reserved.

Cryo-electron microscopy and three-dimensional reconstructions of hepatitis C virus particles

International Nuclear Information System (INIS)

Yu Xuekui; Qiao Ming; Atanasov, Ivo; Hu Zongyi; Kato, Takanobu; Liang, T. Jake; Zhou, Z. Hong

2007-01-01

The structural details of hepatitis C virus (HCV) have been elusive because of the lack of a robust tissue culture system for producing an adequate amount of virions from infectious sources for in-depth three-dimensional (3D) structural analysis. Using both negative-stain and cryo-electron microscopy (cryoEM), we show that HCV virions isolated from cell culture have a rather uniform size of 500 A in diameter and that recombinantly expressed HCV-like particles (HCV-LPs) have similar morphologic, biophysical and antigenic features in spite of the varying sizes of the particles. 3D reconstructions were obtained from HCV-LPs with the same size as the HCV virions in the presence and absence of monoclonal antibodies bound to the E1 glycoprotein. The 3D reconstruction of HCV-LP reveals a multilayered architecture, with smooth outer-layer densities arranged in a 'fishbone' configuration. Reconstruction of the particles in complex with anti-E1 antibodies shows that sites of the E1 epitope are exposed and surround the 5-, 3- and 2-fold axes. The binding pattern of the anti-E1 antibody and the fitting of the structure of the dengue virus E glycoprotein into our 3D reconstructions further suggest that the HCV-LP E1 and E2 proteins form a tetramer (or dimer of heterodimers) that corresponds morphologically and functionally to the flavivirus E homodimer. This first 3D structural analysis of HCV particles offers important insights into the elusive mechanisms of HCV assembly and maturation

Label-free visualization of ultrastructural features of artificial synapses via cryo-EM.

Science.gov (United States)

Gopalakrishnan, Gopakumar; Yam, Patricia T; Madwar, Carolin; Bostina, Mihnea; Rouiller, Isabelle; Colman, David R; Lennox, R Bruce

2011-12-21

The ultrastructural details of presynapses formed between artificial substrates of submicrometer silica beads and hippocampal neurons are visualized via cryo-electron microscopy (cryo-EM). The silica beads are derivatized by poly-d-lysine or lipid bilayers. Molecular features known to exist at presynapses are clearly present at these artificial synapses, as visualized by cryo-EM. Key synaptic features such as the membrane contact area at synaptic junctions, the presynaptic bouton containing presynaptic vesicles, as well as microtubular structures can be identified. This is the first report of the direct, label-free observation of ultrastructural details of artificial synapses.

Effects of residual aberrations explored on single-walled carbon nanotubes

International Nuclear Information System (INIS)

Biskupek, Johannes; Hartel, Peter; Haider, Maximilian; Kaiser, Ute

2012-01-01

The effects of geometric residual aberrations such as coma B 2 and two-fold astigmatism A 1 on the contrast in aberration corrected high resolution transmission electron microscopy (HRTEM) images are investigated on single-walled carbon nanotubes (SWNT). The individual aberrations are adjusted and set up manually using an imaging C S -corrector. We demonstrate how coma B 2 can be recognized by an experienced user directly in the image and how it blurs the contrast. Even with uncorrected (resolution limiting) spherical aberration C S the coma B 2 has to be considered and must be minimized. Limits for a tolerable coma are given. The experiments are confirmed by image simulations. -- Highlights: ► Individual effects of residual aberrations such as B 2 , A 1 , and C S are demonstrated. ► Experimental HRTEM and simulated images of carbon nanotubes are compared. ► A detection limit of 50 nm B 2 in a single HRTEM image is determined.

Index mismatch aberration correction over long working distances using spatial light modulation.

Science.gov (United States)

Gjonaj, Bergin; Johnson, Patrick; Bonn, Mischa; Domke, Katrin F

2012-11-20

For many microscopy applications, millimeters-long free working distances (LWD) are required. However, the high resolution and contrast of LWD objectives operated in air are lost when introducing glass and/or liquid with the sample. We propose to use spatial light modulation to correct for such beam aberrations caused by refractive index mismatches. Focusing a monochromatic laser beam with a 10 mm working distance air objective (50×, 0.5 NA) through air, glass, and water, we manage to restore a sharp, intense focus (FWHMLWD air objectives, extending their usage beyond applications in air.

Electron tomography of cryo-immobilized plant tissue: a novel approach to studying 3D macromolecular architecture of mature plant cell walls in situ.

Directory of Open Access Journals (Sweden)

Purbasha Sarkar

Full Text Available Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (∼ 2 nm, and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF, cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we

Molecular architecture of axonemal microtubule doublets revealedby cryo-electron tomography

Energy Technology Data Exchange (ETDEWEB)

Sui, Haixin; Downing, Kenneth H.

2006-05-22

The axoneme, which forms the core of eukaryotic flagella and cilia, is one of the largest macromolecular machines with a structure that is largely conserved from protists to mammals. Microtubule doublets are structural components of axonemes containing a number of proteins besides tubulin, and are usually found in arrays of nine doublets arranged around two singlet microtubules. Coordinated sliding of adjacent doublets, which involves a host of other proteins in the axoneme, produces periodic beating movements of the axoneme. We have obtained a 3D density map of intact microtubule doublets using cryo-electron tomography and image averaging. Our map, with a resolution of about 3 nm, provides insights into locations of particular proteins within the doublets and the structural features of the doublets that define their mechanical properties. We identify likely candidates for several of these non-tubulin components of the doublets. This work offers novel insight on how tubulin protofilaments and accessory proteins attach together to form the doublets and provides a structural basis for understanding doublet function in axonemes.

Optimized cryo-focused ion beam sample preparation aimed at in situ structural studies of membrane proteins.

Science.gov (United States)

Schaffer, Miroslava; Mahamid, Julia; Engel, Benjamin D; Laugks, Tim; Baumeister, Wolfgang; Plitzko, Jürgen M

2017-02-01

While cryo-electron tomography (cryo-ET) can reveal biological structures in their native state within the cellular environment, it requires the production of high-quality frozen-hydrated sections that are thinner than 300nm. Sample requirements are even more stringent for the visualization of membrane-bound protein complexes within dense cellular regions. Focused ion beam (FIB) sample preparation for transmission electron microscopy (TEM) is a well-established technique in material science, but there are only few examples of biological samples exhibiting sufficient quality for high-resolution in situ investigation by cryo-ET. In this work, we present a comprehensive description of a cryo-sample preparation workflow incorporating additional conductive-coating procedures. These coating steps eliminate the adverse effects of sample charging on imaging with the Volta phase plate, allowing data acquisition with improved contrast. We discuss optimized FIB milling strategies adapted from material science and each critical step required to produce homogeneously thin, non-charging FIB lamellas that make large areas of unperturbed HeLa and Chlamydomonas cells accessible for cryo-ET at molecular resolution. Copyright © 2016 Elsevier Inc. All rights reserved.

Human Skin Barrier Structure and Function Analyzed by Cryo-EM and Molecular Dynamics Simulation.

Science.gov (United States)

Lundborg, Magnus; Narangifard, Ali; Wennberg, Christian L; Lindahl, Erik; Daneholt, Bertil; Norlén, Lars

2018-04-24

In the present study we have analyzed the molecular structure and function of the human skin's permeability barrier using molecular dynamics simulation validated against cryo-electron microscopy data from near native skin. The skin's barrier capacity is located to an intercellular lipid structure embedding the cells of the superficial most layer of skin - the stratum corneum. According to the splayed bilayer model (Iwai et al., 2012) the lipid structure is organized as stacked bilayers of ceramides in a splayed chain conformation with cholesterol associated with the ceramide sphingoid moiety and free fatty acids associated with the ceramide fatty acid moiety. However, knowledge about the lipid structure's detailed molecular organization, and the roles of its different lipid constituents, remains circumstantial. Starting from a molecular dynamics model based on the splayed bilayer model, we have, by stepwise structural and compositional modifications, arrived at a thermodynamically stable molecular dynamics model expressing simulated electron microscopy patterns matching original cryo-electron microscopy patterns from skin extremely closely. Strikingly, the closer the individual molecular dynamics models' lipid composition was to that reported in human stratum corneum, the better was the match between the models' simulated electron microscopy patterns and the original cryo-electron microscopy patterns. Moreover, the closest-matching model's calculated water permeability and thermotropic behaviour were found compatible with that of human skin. The new model may facilitate more advanced physics-based skin permeability predictions of drugs and toxicants. The proposed procedure for molecular dynamics based analysis of cellular cryo-electron microscopy data might be applied to other biomolecular systems. Copyright © 2018. Published by Elsevier Inc.

The cryo-electron microscopy structure of huntingtin

Science.gov (United States)

Guo, Qiang; Bin Huang; Cheng, Jingdong; Seefelder, Manuel; Engler, Tatjana; Pfeifer, Günter; Oeckl, Patrick; Otto, Markus; Moser, Franziska; Maurer, Melanie; Pautsch, Alexander; Baumeister, Wolfgang; Fernández-Busnadiego, Rubén; Kochanek, Stefan

2018-03-01

Huntingtin (HTT) is a large (348 kDa) protein that is essential for embryonic development and is involved in diverse cellular activities such as vesicular transport, endocytosis, autophagy and the regulation of transcription. Although an integrative understanding of the biological functions of HTT is lacking, the large number of identified HTT interactors suggests that it serves as a protein-protein interaction hub. Furthermore, Huntington’s disease is caused by a mutation in the HTT gene, resulting in a pathogenic expansion of a polyglutamine repeat at the amino terminus of HTT. However, only limited structural information regarding HTT is currently available. Here we use cryo-electron microscopy to determine the structure of full-length human HTT in a complex with HTT-associated protein 40 (HAP40; encoded by three F8A genes in humans) to an overall resolution of 4 Å. HTT is largely α-helical and consists of three major domains. The amino- and carboxy-terminal domains contain multiple HEAT (huntingtin, elongation factor 3, protein phosphatase 2A and lipid kinase TOR) repeats arranged in a solenoid fashion. These domains are connected by a smaller bridge domain containing different types of tandem repeats. HAP40 is also largely α-helical and has a tetratricopeptide repeat-like organization. HAP40 binds in a cleft and contacts the three HTT domains by hydrophobic and electrostatic interactions, thereby stabilizing the conformation of HTT. These data rationalize previous biochemical results and pave the way for improved understanding of the diverse cellular functions of HTT.

Heparin binding sites on Ross River virus revealed by electron cryo-microscopy

International Nuclear Information System (INIS)

Zhang Wei; Heil, Marintha; Kuhn, Richard J.; Baker, Timothy S.

2005-01-01

Cell surface glycosaminoglycans play important roles in cell adhesion and viral entry. Laboratory strains of two alphaviruses, Sindbis and Semliki Forest virus, have been shown to utilize heparan sulfate as an attachment receptor, whereas Ross River virus (RRV) does not significantly interact with it. However, a single amino acid substitution at residue 218 in the RRV E2 glycoprotein adapts the virus to heparan sulfate binding and expands the host range of the virus into chicken embryo fibroblasts. Structures of the RRV mutant, E2 N218R, and its complex with heparin were determined through the use of electron cryo-microscopy and image reconstruction methods. Heparin was found to bind at the distal end of the RRV spikes, in a region of the E2 glycoprotein that has been previously implicated in cell-receptor recognition and antibody binding

Calculations of time-of-flight aberrations in practical electrostatic electron lenses using the differential algebraic method

International Nuclear Information System (INIS)

Kang, Yongfeng; Zhao, Jingyi; Tang, Tiantong

2013-01-01

The high order time-of-flight (TOF) aberrations in a practical electrostatic electron lens are calculated using the differential algebraic (DA) method. The electrostatic fields of the electrostatic lens, which are calculated by the FEM methods, are in the form of discrete arrays. Thus, the proposed DA method is applicable for engineering designs, and programs are written to compute up to fifth order TOF aberrations of practical electrostatic electron lenses. An example is given, and TOF aberrations up to the fifth order are calculated. It is proven that the numerical results for the electrostatic fields in the form of discrete arrays have a good accuracy compared with the theoretical solutions. The accuracy is limited only by the accuracy of the numerical computation of the fields and the numerical computation algorithms for interpolation and integration. Finally, a practical electrostatic electron lens is analysed and discussed as an example.

Retrofit implementation of Zernike phase plate imaging for cryo-TEM.

Science.gov (United States)

Marko, Michael; Leith, Ardean; Hsieh, Chyongere; Danev, Radostin

2011-05-01

In-focus phase-plate imaging is particularly beneficial for cryo-TEM because it offers a substantial overall increase in image contrast, without an electron dose penalty, and it simplifies image interpretation. We show how phase-plate cryo-TEM can be implemented with an appropriate existing TEM, and provide a basic practical introduction to use of thin-film (carbon) phase plates. We point out potential pitfalls of phase-plate operation, and discuss solutions. We provide information on evaluating a particular TEM for its suitability. Copyright © 2011 Elsevier Inc. All rights reserved.

Efficient estimation of three-dimensional covariance and its application in the analysis of heterogeneous samples in cryo-electron microscopy.

Science.gov (United States)

Liao, Hstau Y; Hashem, Yaser; Frank, Joachim

2015-06-02

Single-particle cryogenic electron microscopy (cryo-EM) is a powerful tool for the study of macromolecular structures at high resolution. Classification allows multiple structural states to be extracted and reconstructed from the same sample. One classification approach is via the covariance matrix, which captures the correlation between every pair of voxels. Earlier approaches employ computing-intensive resampling and estimate only the eigenvectors of the matrix, which are then used in a separate fast classification step. We propose an iterative scheme to explicitly estimate the covariance matrix in its entirety. In our approach, the flexibility in choosing the solution domain allows us to examine a part of the molecule in greater detail. Three-dimensional covariance maps obtained in this way from experimental data (cryo-EM images of the eukaryotic pre-initiation complex) prove to be in excellent agreement with conclusions derived by using traditional approaches, revealing in addition the interdependencies of ligand bindings and structural changes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Robust w-Estimators for Cryo-EM Class Means

Science.gov (United States)

Huang, Chenxi; Tagare, Hemant D.

2016-01-01

A critical step in cryogenic electron microscopy (cryo-EM) image analysis is to calculate the average of all images aligned to a projection direction. This average, called the “class mean”, improves the signal-to-noise ratio in single particle reconstruction (SPR). The averaging step is often compromised because of outlier images of ice, contaminants, and particle fragments. Outlier detection and rejection in the majority of current cryo-EM methods is done using cross-correlation with a manually determined threshold. Empirical assessment shows that the performance of these methods is very sensitive to the threshold. This paper proposes an alternative: a “w-estimator” of the average image, which is robust to outliers and which does not use a threshold. Various properties of the estimator, such as consistency and influence function are investigated. An extension of the estimator to images with different contrast transfer functions (CTFs) is also provided. Experiments with simulated and real cryo-EM images show that the proposed estimator performs quite well in the presence of outliers. PMID:26841397

CryoSat Mission over Polar Region: Data quality status and product evolutions

Science.gov (United States)

Bouffard, J.; Parrinello, T.; Féménias, P.; Fornari, M.; Scagliola, M.; Baker, S.; Brockley, D.; Mannan, R.; Hall, A.; Webb, E.; Garcia-Mondéjar, A.; Roca, M.; Mantovani, P. L.

2015-12-01

Over the past 20 years, satellite radar altimetry has shown its ability to revolutionize our understanding of the ocean and climate. These advances were mainly limited to ice-free regions, leaving aside large portions of Polar Regions. Launched in 2010, the polar-orbiting CryoSat Satellite was designed to measure the changes in the thickness of polar sea ice and the elevation of the ice sheets and mountain glaciers. To reach this goal, the CryoSat products have to meet the highest performance, through constant improvements of the associated Instrument Processing Facility. Since April 2015, the CryoSat ice products are generated with the Baseline C; which represents a major processor upgrade. Several improvements have been implemented belong this new Baseline, such as SAR retracker optimized for Freeboard retrieval and a coarse slant correction, which is applied directly on the stack data in conjunction with the window delay alignment. The resulting waveforms show more power and the trailing edge is modified, leading to improved L2 geophysical parameters. This paper provides an overview of the CryoSat data characteristics, assessment and exploitation over Polar Regions. In this respect, new science-oriented diagnostics have been implemented to thoroughly understand the signatures within the altimeter signals over sea-ice and land ice areas, to validate the data and therefore propose potential way of improvements for next CryoSat processing Baselines.

Effect of shroud material on the spherical aberration in electromagnetic focusing lens used in electron beam welding machines

International Nuclear Information System (INIS)

Saha, Srijit Kumar; Gupta, Sachin; Kandaswamy, E.

2015-01-01

Beam Power density on the target (typically 10"5 -10"6 W/cm"2 ) plays a major role in attaining good weld quality in electron beam welding. Spherical aberration in the electromagnetic focusing lenses places a limitation in attaining the required power density on the target. Conventionally, iron or low carbon steel core are being used as a shroud material in the electromagnetic lenses. The practical difficulty faced in the long term performance of these lenses has initiated a systematic study for various shroud materials and the effect on spherical aberration limited spot size. The particle trajectories were simulated with different magnetic materials, using commercial software. The spherical aberration was found to be the lowest in the air core lens. The possibility of using an aircore electromagnetic focusing lens in electron beam machines is discussed in this paper. The beam power density is limited by various factors such as spherical aberration, space charge aberrations, gun alignment and power source parameters. (author)

A comparative analysis of the cryo-compression and cryo-adsorption hydrogen storage methods

Energy Technology Data Exchange (ETDEWEB)

Petitpas, G [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Benard, P [Universite du Quebec a Trois-Rivieres (Canada); Klebanoff, L E [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Xiao, J [Universite du Quebec a Trois-Rivieres (Canada); Aceves, S M [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

2014-07-01

While conventional low-pressure LH₂ dewars have existed for decades, advanced methods of cryogenic hydrogen storage have recently been developed. These advanced methods are cryo-compression and cryo-adsorption hydrogen storage, which operate best in the temperature range 30–100 K. We present a comparative analysis of both approaches for cryogenic hydrogen storage, examining how pressure and/or sorbent materials are used to effectively increase onboard H₂ density and dormancy. We start by reviewing some basic aspects of LH₂ properties and conventional means of storing it. From there we describe the cryo-compression and cryo-adsorption hydrogen storage methods, and then explore the relationship between them, clarifying the materials science and physics of the two approaches in trying to solve the same hydrogen storage task (~5–8 kg H₂, typical of light duty vehicles). Assuming that the balance of plant and the available volume for the storage system in the vehicle are identical for both approaches, the comparison focuses on how the respective storage capacities, vessel weight and dormancy vary as a function of temperature, pressure and type of cryo-adsorption material (especially, powder MOF-5 and MIL-101). By performing a comparative analysis, we clarify the science of each approach individually, identify the regimes where the attributes of each can be maximized, elucidate the properties of these systems during refueling, and probe the possible benefits of a combined “hybrid” system with both cryo-adsorption and cryo-compression phenomena operating at the same time. In addition the relationships found between onboard H₂ capacity, pressure vessel and/or sorbent mass and dormancy as a function of rated pressure, type of sorbent material and fueling conditions are useful as general designing guidelines in future engineering efforts using these two hydrogen storage approaches.

Quantum ballistic transistor and low noise HEMT for cryo-electronics lower than 4.2 K; Transistor balistique quantique et HEMT bas-bruit pour la cryoelectronique inferieure a 4.2 K

Energy Technology Data Exchange (ETDEWEB)

Gremion, E

2008-01-15

Next generations of cryo-detectors, widely used in physics of particles and physics of universe, will need in the future high-performance cryo-electronics less noisy and closer to the detector. Within this context, this work investigates properties of two dimensional electron gas GaAlAs/GaAs by studying two components, quantum point contact (QPC) and high electron mobility transistor (HEMT). Thanks to quantized conductance steps in QPC, we have realized a quantum ballistic transistor (voltage gain higher than 1), a new component useful for cryo-electronics thanks to its operating temperature and weak power consumption (about 1 nW). Moreover, the very low capacity of this component leads to promising performances for multiplexing low temperature bolometer dedicated to millimetric astronomy. The second study focused on HEMT with very high quality 2DEG. At 4.2 K, a voltage gain higher than 20 can be obtained with a very low power dissipation of less than 100 {mu}W. Under the above experimental conditions, an equivalent input voltage noise of 1.2 nV/{radical}(Hz) at 1 kHz and 0.12 nV/{radical}(Hz) at 100 kHz has been reached. According to the Hooge formula, these noise performances are get by increasing gate capacity estimated to 60 pF. (author)

CryoSat Land Ice Product Validation within the CryoVal-LI project

DEFF Research Database (Denmark)

Sørensen, Louise Sandberg; Baker, Steven; Csatho, Bea

The main objective of the ESA funded CryoVal-LI project has been to identify and quantify the error sources for the CryoSat-2 mission over land ice. This has been undertaken through the careful documentation of the possible error sources, the identification of suitable validation sites...

Calibration And Validation Of CryoSat-2 Low Resolution Mode Data

Science.gov (United States)

Naeije, M.; Schrama, E.; Scharroo, R.

2011-02-01

Running ahead of the continuously growing need for operational use of sea level products, TUDelft started off the Radar Altimeter Database System RADS many years ago. This system attends to a global international sea- level service. It supports, on one hand, science, like studies on ocean circulation, El Nio, sea level change, and ice topography, and on the other hand (offshore) operations, like delivery of ocean current information, wind and wave statistics, ice detection and ice classification. At present, the database is used by a large scientific community throughout the world, and is daily maintained and developed by Altimetrics LLC, TUDelft and NOAA. It contains all historic altimeter data, and now has to be up- dated with the data from ESAs ice mission CryoSat-2, which was launched successfully in April 2010. These new data are important to augment the data set and by that to improve the estimates of sea level change and its contributors. For this the data have to be validated and calibrated, necessary corrections added and improved (including modelling of corrections that are not directly available from the CryoSat-2 platform), and the orbit ac- curacy verified and if possible the orbits brushed up. Subsequently, value-added ocean and ice products need to be developed in synergy with all the other satellite altimeter data. During the commissioning phase we primarily looked at the sanity of the available level-1b and level-2 Low Resolution Mode (LRM) data. Here, for the 2011 CryoSat Validation Workshop, we present the results of our calibration and validation of LRM L2 data by internal comparison of CryoSat-2 and external comparison with other satellites. We have established a range bias of 3.77 (measurement range too long) and a timing bias of 8.2ms (measurement range too late).

Dose-response relationship for chromosomal aberrations induced in human lymphocytes by 18 MeV electron beam irradiation

International Nuclear Information System (INIS)

Lashin, E.A.; Elaasar, E.M.; Moustafa, H.F.; Bakir, Y.Y.; Al Zenki, S.D.

1990-01-01

Dose response curves for lymphocyte chromosome aberration frequencies using X- and gamma radiation became an important and reliable indicator as biological dosimeter especially in radiation accidents and occupational over exposures. Nowadays electron beam therapy is frequently used for their advantages in cases of tumours under or near to the body surface. Dose-response curves for these electron beams are rarely published. Human peripheral blood lymphocytes were in vitro irradiated with various low and high doses (0.1 Gy to 4.9 Gy) of 18 MeV electron beams to utilize such a dose-response curve using chromosomal aberration frequencies as a biological indicator. Then we compared the biological curve with physically obtained curves normally used in planning for radiotherapy treatment. It is interesting to find a significant difference between both of them. The biological curve is generally higher in value and the aberrations induced by 93% of a dose is significantly higher and deeper in site than those aberrations induced by the 100% dose calculated physically. If the above observation is confirmed by detailed studies, it would be of importance to the radiotherapist to plan for isodose curves according to biological determinations. (author)

CryoSat Level1b SAR/SARin: quality improvements towards BaselineC

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; Bouzinac, Catherine; Tagliani, Nicolas; Parrinello, Tommaso

2014-05-01

CryoSat was launched on the 8th April 2010 and it is the first European ice mission dedicated to monitoring precise changes in the thickness of polar ice sheets and floating sea ice over a 3-year period. Cryosat carries an innovative radar altimeter called the Synthetic Aperture Interferometric Altimeter (SIRAL), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach a significantly improved along track resolution with respect to traditional pulse-width limited altimeters. CryoSat is the first altimetry mission operating in SAR mode and continuous improvement in the Level1 Instrument Processing Facility (IPF1) are being identified, tested and validated in order to improve the quality of the Level1b products. Towards the release of the BaselineC of the CryoSat Level1b SAR/SARin products, that is expected during 2014, several improvements have been identified: • a datation bias of about -0.5195 ms will be corrected • a range bias of about -0.6730 m will be corrected • the waveform length in the Level1b product will be doubled with respect to BaselineB • improved processing for 1Hz echoes to have sharper waveforms • surface sample stack weighting to filter out the single look echoes acquired at highest look angle, that results in a sharpening of the 20Hz waveforms This poster details the main improvements that are foreseen to be included in the CryoSat Level1b SAR/SARin products in BaselineC.

CryoSat SAR/SARin Level1b products: assessment of BaselineC and improvements towards BaselineD

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; Bouffard, Jerome; Parrinello, Tommaso

2017-04-01

CryoSat was launched on the 8th April 2010 and is the first European ice mission dedicated to the monitoring of precise changes in the thickness of polar ice sheets and floating sea ice. Cryosat carries an innovative radar altimeter called the Synthetic Aperture Interferometric Altimeter (SIRAL), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach a significantly improved along track resolution with respect to traditional pulse-width limited altimeters. CryoSat is the first altimetry mission operating in SAR mode and continuous improvements in the Level1 Instrument Processing Facility (IPF1) are being identified, tested and validated in order to improve the quality of the Level1b products. The current IPF, Baseline C, was released in operation in April 2015 and the second CryoSat reprocessing campaign was jointly initiated, taking benefit of the upgrade implemented in the IPF1 processing chain but also of some specific configurations for the calibration corrections. In particular, the CryoSat Level1b BaselineC products generated in the framework of the second reprocessing campaign include refined information for what concerns the mispointing angles and the calibration corrections. This poster will thus detail thus the evolutions that are currently planned for the CryoSat BaselineD SAR/SARin Level1b products and the corresponding quality improvements that are expected.

Cryo-CMOS Circuits and Systems for Quantum Computing Applications

NARCIS (Netherlands)

Patra, B; Incandela, R.M.; van Dijk, J.P.G.; Homulle, H.A.R.; Song, Lin; Shahmohammadi, M.; Staszewski, R.B.; Vladimirescu, A.; Babaie, M.; Sebastiano, F.; Charbon, E.E.E.

2018-01-01

A fault-tolerant quantum computer with millions of quantum bits (qubits) requires massive yet very precise control electronics for the manipulation and readout of individual qubits. CMOS operating at cryogenic temperatures down to 4 K (cryo-CMOS) allows for closer system integration, thus promising

Effect of aberrations in human eye on contrast sensitivity function

Science.gov (United States)

Quan, Wei; Wang, Feng-lin; Wang, Zhao-qi

2011-06-01

The quantitative analysis of the effect of aberrations in human eye on vision has important clinical value in the correction of aberrations. The wave-front aberrations of human eyes were measured with the Hartmann-Shack wave-front sensor and modulation transfer function (MTF) was computed from the wave-front aberrations. Contrast sensitivity function (CSF) was obtained from MTF and the retinal aerial image modulation (AIM). It is shown that the 2nd, 3rd, 4th, 5th, 6th Zernike aberrations deteriorate contrast sensitivity function. When the 2nd, 3rd, 4th, 5th, 6th Zernike aberrations are corrected high contrast sensitivity function can be obtained.

CryoSat-2: Post launch performance of SIRAL-2 and its calibration/validation

Science.gov (United States)

Cullen, Robert; Francis, Richard; Davidson, Malcolm; Wingham, Duncan

2010-05-01

1. INTRODUCTION The main payload of CryoSat-2 [1], SIRAL (Synthetic interferometric radar altimeter), is a Ku band pulse-width limited radar altimeter which transmits pulses at a high pulse repetition frequency thus making received echoes phase coherent and suitable for azimuth processing [2]. The azimuth processing in conjunction with correction for slant range improves along track resolution to about 250 meters which is a significant improvement over traditional pulse-width limited systems such as Envisat RA-2, [3]. CryoSat-2 will be launched on 25th February 2010 and this paper describes the pre and post launch measures of CryoSat/SIRAL performance and the status of mission validation planning. 2. SIRAL PERFORMANCE: INTERNAL AND EXTERNAL CALIBRATION Phase coherent pulse-width limited radar altimeters such as SIRAL-2 pose a new challenge when considering a strategy for calibration. Along with the need to generate the well understood corrections for transfer function amplitude with respect to frequency, gain and instrument path delay there is also a need to provide corrections for transfer function phase with respect to frequency and AGC setting, phase variation across bursts of pulses. Furthermore, since some components of these radars are temperature sensitive one needs to be careful when the deciding how often calibrations are performed whilst not impacting mission performance. Several internal calibration ground processors have been developed to model imperfections within the CryoSat-2 radar altimeter (SIRAL-2) hardware and reduce their effect from the science data stream via the use of calibration correction auxiliary products within the ground segment. We present the methods and results used to model and remove imperfections and describe the baseline for usage of SIRAL-2 calibration modes during the commissioning phase and the operational exploitation phases of the mission. Additionally we present early results derived from external calibration of SIRAL via

Mask-induced aberration in EUV lithography

Science.gov (United States)

Nakajima, Yumi; Sato, Takashi; Inanami, Ryoichi; Nakasugi, Tetsuro; Higashiki, Tatsuhiko

2009-04-01

We estimated aberrations using Zernike sensitivity analysis. We found the difference of the tolerated aberration with line direction for illumination. The tolerated aberration of perpendicular line for illumination is much smaller than that of parallel line. We consider this difference to be attributable to the mask 3D effect. We call it mask-induced aberration. In the case of the perpendicular line for illumination, there was a difference in CD between right line and left line without aberration. In this report, we discuss the possibility of pattern formation in NA 0.25 generation EUV lithography tool. In perpendicular pattern for EUV light, the dominant part of aberration is mask-induced aberration. In EUV lithography, pattern correction based on the mask topography effect will be more important.

Performance evaluation of spatial compounding in the presence of aberration and adaptive imaging

Science.gov (United States)

Dahl, Jeremy J.; Guenther, Drake; Trahey, Gregg E.

2003-05-01

Spatial compounding has been used for years to reduce speckle in ultrasonic images and to resolve anatomical features hidden behind the grainy appearance of speckle. Adaptive imaging restores image contrast and resolution by compensating for beamforming errors caused by tissue-induced phase errors. Spatial compounding represents a form of incoherent imaging, whereas adaptive imaging attempts to maintain a coherent, diffraction-limited aperture in the presence of aberration. Using a Siemens Antares scanner, we acquired single channel RF data on a commercially available 1-D probe. Individual channel RF data was acquired on a cyst phantom in the presence of a near field electronic phase screen. Simulated data was also acquired for both a 1-D and a custom built 8x96, 1.75-D probe (Tetrad Corp.). The data was compounded using a receive spatial compounding algorithm; a widely used algorithm because it takes advantage of parallel beamforming to avoid reductions in frame rate. Phase correction was also performed by using a least mean squares algorithm to estimate the arrival time errors. We present simulation and experimental data comparing the performance of spatial compounding to phase correction in contrast and resolution tasks. We evaluate spatial compounding and phase correction, and combinations of the two methods, under varying aperture sizes, aperture overlaps, and aberrator strength to examine the optimum configuration and conditions in which spatial compounding will provide a similar or better result than adaptive imaging. We find that, in general, phase correction is hindered at high aberration strengths and spatial frequencies, whereas spatial compounding is helped by these aberrators.

Breaking Symmetry in Viral Icosahedral Capsids as Seen through the Lenses of X-ray Crystallography and Cryo-Electron Microscopy

Directory of Open Access Journals (Sweden)

Kristin N. Parent

2018-02-01

Full Text Available The majority of viruses on Earth form capsids built by multiple copies of one or more types of a coat protein arranged with 532 symmetry, generating an icosahedral shell. This highly repetitive structure is ideal to closely pack identical protein subunits and to enclose the nucleic acid genomes. However, the icosahedral capsid is not merely a passive cage but undergoes dynamic events to promote packaging, maturation and the transfer of the viral genome into the host. These essential processes are often mediated by proteinaceous complexes that interrupt the shell’s icosahedral symmetry, providing a gateway through the capsid. In this review, we take an inventory of molecular structures observed either internally, or at the 5-fold vertices of icosahedral DNA viruses that infect bacteria, archea and eukaryotes. Taking advantage of the recent revolution in cryo-electron microscopy (cryo-EM and building upon a wealth of crystallographic structures of individual components, we review the design principles of non-icosahedral structural components that interrupt icosahedral symmetry and discuss how these macromolecules play vital roles in genome packaging, ejection and host receptor-binding.

Seeing with electrons

International Nuclear Information System (INIS)

Nellist, P.

2006-01-01

Commercially available lens correctors are extending the reach of electron microscopes to unprecedented atomic scales, as Peter Nellist describes. The electron microscope was invented in 1933 and is based on the principle that electrons have a wavelength that is inversely proportional to their momentum. There are two basic types: transmission electron microscopes and scanning electron microscopes, plus a hybrid of the two. The lenses in an electron microscope are provided by electromagnetic fields, but they suffer from spherical aberration. The addition of octupole and quadrupole corrector fields has improved the resolution of the electron microscope to better than 0.1 nm in the last decade. The next step is to correct for chromatic aberration, after which the resolution of the microscope will probably be limited by the size of the atom itself. (U.K.)

Development of a dielectric barrier discharge (DBD) cryo-microplasma: generation and diagnostics

International Nuclear Information System (INIS)

Ishihara, Daisuke; Noma, Yuri; Stauss, Sven; Sai, Masaki; Tomai, Takaaki; Terashima, Kazuo

2008-01-01

We developed a cryo-microplasma, which can continuously control gas temperature below room temperature and below the freezing point of water. To develop the cryo-microplasma, we first developed an atmospheric-pressure low-temperature microplasma that can suppress the increase in its gas temperature. Helium gas was employed, which was generated in open air. The average estimated electron density and temperature were 10 8 -10 9 cm -3 and 4-5 eV, respectively, independent of the applied voltage. Then, helium gas, which was the working gas of the atmospheric-pressure low-temperature microplasma, was cooled by liquid nitrogen to generate an atmospheric-pressure cryo-microplasma in open air. We observed the generation of frost around the quartz tube in which the plasma was generated and an increase in atomic oxygen emission by optical emission spectroscopy. Finally, to avoid the generation of frost, a cryo-microplasma was generated in a reactor chamber separated from open air. Helium, nitrogen and oxygen were employed as working gases. Using thermocouples and by estimation from the nitrogen rotational temperature, we verified that the gas temperature of the cryo-microplasma was much lower (T g ∼ 180-300 K) than that of the conventional atmospheric-pressure low-temperature plasma (above 300 K).

Development of a dielectric barrier discharge (DBD) cryo-microplasma: generation and diagnostics

Science.gov (United States)

Ishihara, Daisuke; Noma, Yuri; Stauss, Sven; Sai, Masaki; Tomai, Takaaki; Terashima, Kazuo

2008-08-01

We developed a cryo-microplasma, which can continuously control gas temperature below room temperature and below the freezing point of water. To develop the cryo-microplasma, we first developed an atmospheric-pressure low-temperature microplasma that can suppress the increase in its gas temperature. Helium gas was employed, which was generated in open air. The average estimated electron density and temperature were 108-109 cm-3 and 4-5 eV, respectively, independent of the applied voltage. Then, helium gas, which was the working gas of the atmospheric-pressure low-temperature microplasma, was cooled by liquid nitrogen to generate an atmospheric-pressure cryo-microplasma in open air. We observed the generation of frost around the quartz tube in which the plasma was generated and an increase in atomic oxygen emission by optical emission spectroscopy. Finally, to avoid the generation of frost, a cryo-microplasma was generated in a reactor chamber separated from open air. Helium, nitrogen and oxygen were employed as working gases. Using thermocouples and by estimation from the nitrogen rotational temperature, we verified that the gas temperature of the cryo-microplasma was much lower (Tg ≈ 180-300 K) than that of the conventional atmospheric-pressure low-temperature plasma (above 300 K).

Cryo-EM studies of the structure and dynamics of vacuolar-type ATPases

Science.gov (United States)

Mazhab-Jafari, Mohammad T.; Rubinstein, John L.

2016-01-01

Electron cryomicroscopy (cryo-EM) has significantly advanced our understanding of molecular structure in biology. Recent innovations in both hardware and software have made cryo-EM a viable alternative for targets that are not amenable to x-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. Cryo-EM has even become the method of choice in some situations where x-ray crystallography and NMR spectroscopy are possible but where cryo-EM can determine structures at higher resolution or with less time or effort. Rotary adenosine triphosphatases (ATPases) are crucial to the maintenance of cellular homeostasis. These enzymes couple the synthesis or hydrolysis of adenosine triphosphate to the use or production of a transmembrane electrochemical ion gradient, respectively. However, the membrane-embedded nature and conformational heterogeneity of intact rotary ATPases have prevented their high-resolution structural analysis to date. Recent application of cryo-EM methods to the different types of rotary ATPase has led to sudden advances in understanding the structure and function of these enzymes, revealing significant conformational heterogeneity and characteristic transmembrane α helices that are highly tilted with respect to the membrane. In this Review, we will discuss what has been learned recently about rotary ATPase structure and function, with a particular focus on the vacuolar-type ATPases. PMID:27532044

Effects of ocular aberrations on contrast detection in noise.

Science.gov (United States)

Liang, Bo; Liu, Rong; Dai, Yun; Zhou, Jiawei; Zhou, Yifeng; Zhang, Yudong

2012-08-06

We use adaptive optics (AO) techniques to manipulate the ocular aberrations and elucidate the effects of these ocular aberrations on contrast detection in a noisy background. The detectability of sine wave gratings at frequencies of 4, 8, and 16 circles per degree (cpd) was measured in a standard two-interval force-choice staircase procedure against backgrounds of various levels of white noise. The observer's ocular aberrations were either corrected with AO or left uncorrected. In low levels of external noise, contrast detection thresholds are always lowered by AO correction, whereas in high levels of external noise, they are generally elevated by AO correction. Higher levels of external noise are required to make this threshold elevation observable when signal spatial frequencies increase from 4 to 16 cpd. The linear-amplifier-model fit shows that mostly sampling efficiency and equivalent noise both decrease with AO correction. Our findings indicate that ocular aberrations could be beneficial for contrast detection in high-level noises. The implications of these findings are discussed.

Time-resolved GISAXS and cryo-microscopy characterization of block copolymer membrane formation

KAUST Repository

Marques, Debora S.; Dorin, Rachel Mika; Wiesner, Ulrich B.; Smilgies, Detlef Matthias; Behzad, Ali Reza; Vainio, Ulla; Peinemann, Klaus-Viktor; Nunes, Suzana Pereira

2014-01-01

Time-resolved grazing-incidence small-angle X-ray scattering (GISAXS) and cryo-microscopy were used for the first time to understand the pore evolution by copolymer assembly, leading to the formation of isoporous membranes with exceptional porosity and regularity. The formation of copolymer micelle strings in solution (in DMF/DOX/THF and DMF/DOX) was confirmed by cryo field emission scanning electron microscopy (cryo-FESEM) with a distance of 72 nm between centers of micelles placed in different strings. SAXS measurement of block copolymer solutions in DMF/DOX indicated hexagonal assembly with micelle-to-micelle distance of 84-87 nm for 14-20 wt% copolymer solutions. GISAXS in-plane peaks were detected, revealing order close to hexagonal. The d-spacing corresponding to the first peak in this case was 100-130 nm (lattice constant 115-150 nm) for 17 wt% copolymer solutions evaporating up to 100 s. Time-resolved cryo-FESEM showed the formation of incipient pores on the film surface after 4 s copolymer solution casting with distances between void centers of 125 nm. © 2014 Elsevier Ltd. All rights reserved.

Time-resolved GISAXS and cryo-microscopy characterization of block copolymer membrane formation

KAUST Repository

Marques, Debora S.

2014-03-01

Time-resolved grazing-incidence small-angle X-ray scattering (GISAXS) and cryo-microscopy were used for the first time to understand the pore evolution by copolymer assembly, leading to the formation of isoporous membranes with exceptional porosity and regularity. The formation of copolymer micelle strings in solution (in DMF/DOX/THF and DMF/DOX) was confirmed by cryo field emission scanning electron microscopy (cryo-FESEM) with a distance of 72 nm between centers of micelles placed in different strings. SAXS measurement of block copolymer solutions in DMF/DOX indicated hexagonal assembly with micelle-to-micelle distance of 84-87 nm for 14-20 wt% copolymer solutions. GISAXS in-plane peaks were detected, revealing order close to hexagonal. The d-spacing corresponding to the first peak in this case was 100-130 nm (lattice constant 115-150 nm) for 17 wt% copolymer solutions evaporating up to 100 s. Time-resolved cryo-FESEM showed the formation of incipient pores on the film surface after 4 s copolymer solution casting with distances between void centers of 125 nm. © 2014 Elsevier Ltd. All rights reserved.

EMHP: an accurate automated hole masking algorithm for single-particle cryo-EM image processing.

Science.gov (United States)

Berndsen, Zachary; Bowman, Charles; Jang, Haerin; Ward, Andrew B

2017-12-01

The Electron Microscopy Hole Punch (EMHP) is a streamlined suite of tools for quick assessment, sorting and hole masking of electron micrographs. With recent advances in single-particle electron cryo-microscopy (cryo-EM) data processing allowing for the rapid determination of protein structures using a smaller computational footprint, we saw the need for a fast and simple tool for data pre-processing that could run independent of existing high-performance computing (HPC) infrastructures. EMHP provides a data preprocessing platform in a small package that requires minimal python dependencies to function. https://www.bitbucket.org/chazbot/emhp Apache 2.0 License. bowman@scripps.edu. Supplementary data are available at Bioinformatics online. © The Author(s) 2017. Published by Oxford University Press.

Processing a Biological Tissue from Cryo-SEM to Replica

Czech Academy of Sciences Publication Activity Database

Vaškovicová, Naděžda; Hrubanová, Kamila; Krzyžánek, Vladislav

2016-01-01

Roč. 22, S3 (2016), s. 236-237 ISSN 1431-9276 R&D Projects: GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : Cryo-SEM * ACE600 Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.891, year: 2016

Examples of electrostatic electron optics: The Farrand and Elektros microscopes and electron mirrors

International Nuclear Information System (INIS)

Hawkes, P.W.

2012-01-01

The role of Gertrude Rempfer in the design of the Farrand and Elektros microscopes is evoked. The study of electron mirror optics, aberration correction using mirrors and the development of microscopes employing electron mirrors are recapitulated, accompanied by a full bibliography, of earlier publications in particular.

Rooting Out Aberrant Behavior in Training.

Science.gov (United States)

Kokalis, Jerry, Jr.; Paquin, Dave

1989-01-01

Discusses aberrant, or disruptive, behavior in an industrial/business, classroom-based, instructor-led training setting. Three examples of aberrant behavior are described, typical case studies are provided for each, and preventive (long-term) and corrective (on-the-spot) strategies for dealing with the problems are discussed. (LRW)

Pathogen–host reorganization during Chlamydia invasion revealed by cryo-electron tomography

Science.gov (United States)

Nans, Andrea; Saibil, Helen R; Hayward, Richard D

2014-01-01

Invasion of host cells is a key early event during bacterial infection, but the underlying pathogen–host interactions are yet to be fully visualized in three-dimensional detail. We have captured snapshots of the early stages of bacterial-mediated endocytosis in situ by exploiting the small size of chlamydial elementary bodies (EBs) for whole-cell cryo-electron tomography. Chlamydiae are obligate intracellular bacteria that infect eukaryotic cells and cause sexually transmitted infections and trachoma, the leading cause of preventable blindness. We demonstrate that Chlamydia trachomatis LGV2 EBs are intrinsically polarized. One pole is characterized by a tubular inner membrane invagination, while the other exhibits asymmetric periplasmic expansion to accommodate an array of type III secretion systems (T3SSs). Strikingly, EBs orient with their T3SS-containing pole facing target cells, enabling the T3SSs to directly contact the cellular plasma membrane. This contact induces enveloping macropinosomes, actin-rich filopodia and phagocytic cups to zipper tightly around the internalizing bacteria. Once encapsulated into tight early vacuoles, EB polarity and the T3SSs are lost. Our findings reveal previously undescribed structural transitions in both pathogen and host during the initial steps of chlamydial invasion. PMID:24809274

Cryo-electron microscopy structure of the lysosomal calcium-permeable channel TRPML3.

Science.gov (United States)

Hirschi, Marscha; Herzik, Mark A; Wie, Jinhong; Suo, Yang; Borschel, William F; Ren, Dejian; Lander, Gabriel C; Lee, Seok-Yong

2017-10-19

The modulation of ion channel activity by lipids is increasingly recognized as a fundamental component of cellular signalling. The transient receptor potential mucolipin (TRPML) channel family belongs to the TRP superfamily and is composed of three members: TRPML1-TRPML3. TRPMLs are the major Ca 2+ -permeable channels on late endosomes and lysosomes (LEL). They regulate the release of Ca 2+ from organelles, which is important for various physiological processes, including organelle trafficking and fusion. Loss-of-function mutations in the MCOLN1 gene, which encodes TRPML1, cause the neurodegenerative lysosomal storage disorder mucolipidosis type IV, and a gain-of-function mutation (Ala419Pro) in TRPML3 gives rise to the varitint-waddler (Va) mouse phenotype. Notably, TRPML channels are activated by the low-abundance and LEL-enriched signalling lipid phosphatidylinositol-3,5-bisphosphate (PtdIns(3,5)P 2 ), whereas other phosphoinositides such as PtdIns(4,5)P 2 , which is enriched in plasma membranes, inhibit TRPMLs. Conserved basic residues at the N terminus of the channel are important for activation by PtdIns(3,5)P 2 and inhibition by PtdIns(4,5)P 2 . However, owing to a lack of structural information, the mechanism by which TRPML channels recognize PtdIns(3,5)P 2 and increase their Ca 2+ conductance remains unclear. Here we present the cryo-electron microscopy (cryo-EM) structure of a full-length TRPML3 channel from the common marmoset (Callithrix jacchus) at an overall resolution of 2.9 Å. Our structure reveals not only the molecular basis of ion conduction but also the unique architecture of TRPMLs, wherein the voltage sensor-like domain is linked to the pore via a cytosolic domain that we term the mucolipin domain. Combined with functional studies, these data suggest that the mucolipin domain is responsible for PtdIns(3,5)P 2 binding and subsequent channel activation, and that it acts as a 'gating pulley' for lipid-dependent TRPML gating.

CryoSat-2 Validation using CryoVEX 2011-12 Airborne Campaigns

DEFF Research Database (Denmark)

Skourup, Henriette; Forsberg, René; Kildegaard Rose, Stine

Sat-2 by comparison to airborne and ground measurements. This is possible only through a major effort involving a large group of international partners. DTU Space has been involved in the CryoVEx campaigns with airborne activities since 2003. To validate the performance of the CryoSat-2 radar altimeter...... (SIRAL), the aircraft is equipped with an airborne version of the SIRAL altimeter (ASIRAS) together with a laser scanner. The campaigns are focused on five main validation sites: Devon ice cap (Canada), Austfonna ice cap (Svalbard), the EGIG line crossing the Greenland Ice Sheet, as well as the sea ice...... north of Alert and sea ice around Svalbard in the Fram Strait. Selected tracks were planned to match CryoSat-2 passes and a few of them were flown in formation flight with the Alfred Wegener Institute (AWI) Polar-5 carrying an EM-bird. This presentation summarizes the 2011-12 airborne campaigns...

Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy

Directory of Open Access Journals (Sweden)

Johanna L. Höög

2015-11-01

Full Text Available Human ejaculates contain extracellular vesicles (EVs, that to a large extent are considered to originate from the prostate gland, and are often denominated “prostasomes.” These EVs are important for human fertility, for example by promoting sperm motility and by inducing immune tolerance of the female immune system to the spermatozoa. So far, the EVs present in human ejaculate have not been studied in their native state, inside the seminal fluid without prior purification and isolation procedures. Using cryo-electron microscopy and tomography, we performed a comprehensive inventory of human ejaculate EVs. The sample was neither centrifuged, fixed, filtered or sectioned, nor were heavy metals added. Approximately 1,500 extracellular structures were imaged and categorized. The extracellular environment of human ejaculate was found to be diverse, with 5 major subcategories of EVs and 6 subcategories of extracellular membrane compartments, including lamellar bodies. Furthermore, 3 morphological features, including electron density, double membrane bilayers and coated surface, are described in all subcategories. This study reveals that the extracellular environment in human ejaculate is multifaceted. Several novel morphological EV subcategories are identified and clues to their cellular origin may be found in their morphology. This inventory is therefore important for developing future experimental approaches, and to interpret previously published data to understand the role of EVs for human male fertility.

Correção das aberrações oculares nos retratamentos de LASIK personalizado e convencional Correction of ocular aberrations in custom and standard LASIK retreatments

Directory of Open Access Journals (Sweden)

Andréia Peltier Urbano

2009-10-01

Full Text Available OBJETIVO: Comparar a correção das aberrações oculares nos retratamentos de LASIK personalizado e convencional. MÉTODOS: Foi realizado um estudo prospectivo, randomizado, de 74 olhos de 37 pacientes submetidos ao retratamento de LASIK para correção de miopia e astigmatismo. Cada paciente foi submetido ao retratamento de LASIK personalizado Zyoptix (Bausch & Lomb em um olho e convencional Planoscan (Bausch & Lomb no olho contralateral. Foi comparada a correção das aberrações oculares nos retratamentos personalizado e convencional. RESULTADOS: No sexto mês pós-operatório, os olhos submetidos ao retratamento Zyoptix apresentaram diminuição estatisticamente significativa do defocus, astigmatismo, coma, aberração esférica, segunda ordem, terceira ordem, alta ordem e aberrações totais. Os olhos submetidos ao retratamento Planoscan apresentaram diminuição estatisticamente significativa do defocus, segunda ordem e aberrações totais. CONCLUSÕES: O retratamento personalizado foi superior ao retratamento convencional para a correção das aberrações oculares de baixa e alta ordens.PURPOSE: To compare the correction of ocular aberrations between custom and standard LASIK retreatment. METHODS: Prospective, randomized trial with paired eye control of 74 eyes from 37 patients who underwent LASIK retreatment. Each patient underwent retreatment using Zyoptix LASIK (Bausch & Lomb in 1 eye and Planoscan LASIK (Bausch & Lomb in the fellow eye. Correction of ocular aberrations was compared between custom and standard LASIK retreatments. RESULTS: At 6 months, there was a statistically significant reduction in defocus, astigmatism, coma, spherical aberration, second, third, higher-order and total aberration in Zyoptix eyes. There was a statistically significant reduction in defocus, second-order and total aberration in Planoscan eyes. CONCLUSIONS: Custom retreatment was statistically superior than standard retreatment for correction of lower

Deep Tissue Wavefront Estimation for Sensorless Aberration Correction

Directory of Open Access Journals (Sweden)

Ibrahimovic Emina

2015-01-01

Full Text Available The multiple light scattering in biological tissues limits the measurement depth for traditional wavefront sensor. The attenuated ballistic light and the background noise caused by the diffuse light give low signal to noise ratio for wavefront measurement. To overcome this issue, we introduced a wavefront estimation method based on a ray tracing algorithm to overcome this issue. With the knowledge of the refractive index of the medium, the wavefront is estimated by calculating optical path length of rays from the target inside of the samples. This method can provide not only the information of spherical aberration from the refractive-index mismatch between the medium and biological sample but also other aberrations caused by the irregular interface between them. Simulations based on different configurations are demonstrated in this paper.

Structure of the TRPV1 ion channel determined by electron cryo-microscopy.

Science.gov (United States)

Liao, Maofu; Cao, Erhu; Julius, David; Cheng, Yifan

2013-12-05

Transient receptor potential (TRP) channels are sensors for a wide range of cellular and environmental signals, but elucidating how these channels respond to physical and chemical stimuli has been hampered by a lack of detailed structural information. Here we exploit advances in electron cryo-microscopy to determine the structure of a mammalian TRP channel, TRPV1, at 3.4 Å resolution, breaking the side-chain resolution barrier for membrane proteins without crystallization. Like voltage-gated channels, TRPV1 exhibits four-fold symmetry around a central ion pathway formed by transmembrane segments 5-6 (S5-S6) and the intervening pore loop, which is flanked by S1-S4 voltage-sensor-like domains. TRPV1 has a wide extracellular 'mouth' with a short selectivity filter. The conserved 'TRP domain' interacts with the S4-S5 linker, consistent with its contribution to allosteric modulation. Subunit organization is facilitated by interactions among cytoplasmic domains, including amino-terminal ankyrin repeats. These observations provide a structural blueprint for understanding unique aspects of TRP channel function.

DeepPicker: A deep learning approach for fully automated particle picking in cryo-EM.

Science.gov (United States)

Wang, Feng; Gong, Huichao; Liu, Gaochao; Li, Meijing; Yan, Chuangye; Xia, Tian; Li, Xueming; Zeng, Jianyang

2016-09-01

Particle picking is a time-consuming step in single-particle analysis and often requires significant interventions from users, which has become a bottleneck for future automated electron cryo-microscopy (cryo-EM). Here we report a deep learning framework, called DeepPicker, to address this problem and fill the current gaps toward a fully automated cryo-EM pipeline. DeepPicker employs a novel cross-molecule training strategy to capture common features of particles from previously-analyzed micrographs, and thus does not require any human intervention during particle picking. Tests on the recently-published cryo-EM data of three complexes have demonstrated that our deep learning based scheme can successfully accomplish the human-level particle picking process and identify a sufficient number of particles that are comparable to those picked manually by human experts. These results indicate that DeepPicker can provide a practically useful tool to significantly reduce the time and manual effort spent in single-particle analysis and thus greatly facilitate high-resolution cryo-EM structure determination. DeepPicker is released as an open-source program, which can be downloaded from https://github.com/nejyeah/DeepPicker-python. Copyright © 2016 Elsevier Inc. All rights reserved.

Cryo-SEM studies of latex/ceramic nanoparticle coating microstructure development.

Science.gov (United States)

Luo, Hui; Scriven, L E; Francis, Lorraine F

2007-12-15

Cryogenic scanning electron microscopy (cryo-SEM) was used to investigate microstructure development of composite coatings prepared from dispersions of antimony-doped tin oxide (ATO) nanoparticles (approximately 30 nm) or indium tin oxide (ITO) nanoparticles (approximately 40 nm) and latex particles (polydisperse, D(v): approximately 300 nm). Cryo-SEM images of ATO/latex dispersions as-frozen show small clusters of ATO and individual latex particles homogeneously distribute in a frozen water matrix. In contrast, cryo-SEM images of ITO/latex dispersions as-frozen show ITO particles adsorb onto latex particle surfaces. Electrostatic repulsion between negatively charged ATO and negatively charged latex particles stabilizes the ATO/latex dispersion, whereas in ITO/latex dispersion, positively charged ITO particles are attracted onto surfaces of negatively charged latex particles. These results are consistent with calculations of interaction potentials from past research. Cryo-SEM images of frozen and fractured coatings reveal that both ceramic nanoparticles and latex become more concentrated as drying proceeds; larger latex particles consolidate with ceramic nanoparticles in the interstitial spaces. With more drying, compaction flattens the latex-latex particle contacts and shrinks the voids between them. Thus, ceramic nanoparticles are forced to pack closely in the interstitial spaces, forming an interconnected network. Finally, latex particles partially coalesce at their flattened contacts, thereby yielding a coherent coating. The research reveals how nanoparticles segregate and interconnect among latex particles during drying.

On the benefit of the negative-spherical-aberration imaging technique for quantitative HRTEM

International Nuclear Information System (INIS)

Jia, C.L.; Houben, L.; Thust, A.; Barthel, J.

2010-01-01

Employing an aberration corrector in a high-resolution transmission electron microscope, the spherical aberration C S can be tuned to negative values, resulting in a novel imaging technique, which is called the negative C S imaging (NCSI) technique. The image contrast obtained with the NCSI technique is compared quantitatively with the image contrast formed with the traditional positive C S imaging (PCSI) technique. For the case of thin objects negative C S images are superior to positive C S images concerning the magnitude of the obtained contrast, which is due to constructive rather than destructive superposition of fundamental contrast contributions. As a consequence, the image signal obtained with a negative spherical aberration is significantly more robust against noise caused by amorphous surface layers, resulting in a measurement precision of atomic positions which is by a factor of 2-3 better at an identical noise level. The quantitative comparison of the two alternative C S -corrected imaging modes shows that the NCSI mode yields significantly more precise results in quantitative high-resolution transmission electron microscopy of thin objects than the traditional PCSI mode.

Magnitude of speed of sound aberration corrections for ultrasound image guided radiotherapy for prostate and other anatomical sites

International Nuclear Information System (INIS)

Fontanarosa, Davide; Meer, Skadi van der; Bloemen-van Gurp, Esther; Stroian, Gabriela; Verhaegen, Frank

2012-01-01

Purpose: The purpose of this work is to assess the magnitude of speed of sound (SOS) aberrations in three-dimensional ultrasound (US) imaging systems in image guided radiotherapy. The discrepancy between the fixed SOS value of 1540 m/s assumed by US systems in human soft tissues and its actual nonhomogeneous distribution in patients produces small but systematic errors of up to a few millimeters in the positions of scanned structures. Methods: A correction, provided by a previously published density-based algorithm, was applied to a set of five prostate, five liver, and five breast cancer patients. The shifts of the centroids of target structures and the change in shape were evaluated. Results: After the correction the prostate cases showed shifts up to 3.6 mm toward the US probe, which may explain largely the reported positioning discrepancies in the literature on US systems versus other imaging modalities. Liver cases showed the largest changes in volume of the organ, up to almost 9%, and shifts of the centroids up to more than 6 mm either away or toward the US probe. Breast images showed systematic small shifts of the centroids toward the US probe with a maximum magnitude of 1.3 mm. Conclusions: The applied correction in prostate and liver cancer patients shows positioning errors of several mm due to SOS aberration; the errors are smaller in breast cancer cases, but possibly becoming more important when breast tissue thickness increases.

Challenges and opportunities in the high-resolution cryo-EM visualization of microtubules and their binding partners.

Science.gov (United States)

Nogales, Eva; Kellogg, Elizabeth H

2017-10-01

As non-crystallizable polymers, microtubules have been the target of cryo-electron microscopy (cryo-EM) studies since the technique was first established. Over the years, image processing strategies have been developed that take care of the unique, pseudo-helical symmetry of the microtubule. With recent progress in data quality and data processing, cryo-EM reconstructions are now reaching resolutions that allow the generation of atomic models of microtubules and the factors that bind them. These include cellular partners that contribute to microtubule cellular functions, or small ligands that interfere with those functions in the treatment of cancer. The stage is set to generate a family portrait for all identified microtubule interacting proteins and to use cryo-EM as a drug development tool in the targeting of tubulin. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Initial assessment of CryoSat-2 Performance.

Science.gov (United States)

Wingham, D.; Galin, N.; Ridout, A.; Cullen, R.; Giles, K. A.; Laxon, S. W.

2011-12-01

Following the launch of CryoSat-2 in April 2010, we have examined the performance of the CryoSat-2 SAR Interferometer over the continental ice sheets of Antarctica and Greenland, the Artic Ocean, and, for the purposes of calibration, over the oceans. Our aim has been to provide confirmation of the engineering performance of the radar interferometer, and to provide an initial geophysical validation of the resulting elevation measurements. We have confirmed the engineering performance at system level of the interferometer through performing a sequence of satellite rolls over the oceans, which provide a surface of known behavior and surface gradient. The activity has identified some errors in the SARIN L1b data products presently issued by ESA. Once corrected, the ocean calibration has demonstrated that the interferometer measures across-track surface slopes with a precision of 25 micro-radians and an accuracy of 10 micro-radians, which may be compared with a pre-launch estimation of 100 micro-radians; in short, the engineering performance greatly its the specification. The elevation measurement over the ice sheets combines the interferometer measurement of across track slope with the range measurement deduced from the SAR echoes. We have examined the performance of the range estimation, and determined the range precision to be 19 cm RMS at 20 Hz. We have examined the retrieval of the phase information over the ice sheets, and found the phase estimates to be robust and little affected by the uncertain ice sheet topography. Based on the calibration of the interferometer, the contribution of the across track slope error is, at 0.4 mm, negligible. While the quantity of data available to us that contains the corrections identified by the interferometer is limited, we have been able to confirm the range precision values from a limited cross-over analysis. Over marine sea ice, we have verified the discrimination of sea ice and ocean lead returns using contemporaneous SAR

CryoVex 2008

DEFF Research Database (Denmark)

Hvidegaard, Sine Munk; Forsberg, René; Skourup, Henriette

2009-01-01

Air Greenland. The main purpose was to collect coincident ASIRAS and laser elevation data form validation sites on land and sea ice and in addition offer logistical support to ground teams. Overflights of corner reflectors were done at main validation sites in order to calibrate the ASIRAS data....... The datasets from this campaign will be important for understanding the CryoSat-2 radar signals. The airborne part of CryoVEx 2008 was successfully carried out between April 15 and May 8 and the datasets have been stored and secured at DTU Space and Alfred Wegener Institute (AWI). Afterwards extensive data...

Electron cryo-tomographic structure of cystovirus phi 12.

Science.gov (United States)

Hu, Guo-Bin; Wei, Hui; Rice, William J; Stokes, David L; Gottlieb, Paul

2008-03-01

Bacteriophage phi 12 is a member of the Cystoviridae virus family and contains a genome consisting of three segments of double-stranded RNA (dsRNA). This virus family contains eight identified members, of which four have been classified in regard to their complete genomic sequence and encoded viral proteins. A phospholipid envelope that contains the integral proteins P6, P9, P10, and P13 surrounds the viral particles. In species phi 6, host infection requires binding of a multimeric P3 complex to type IV pili. In species varphi8, phi 12, and phi 13, the attachment apparatus is a heteromeric protein assembly that utilizes the rough lipopolysaccharide (rlps) as a receptor. In phi 8 the protein components are designated P3a and P3b while in species phi 12 proteins P3a and P3c have been identified in the complex. The phospholipid envelope of the cystoviruses surrounds a nucleocapsid (NC) composed of two shells. The outer shell is composed of protein P8 with a T=13 icosahedral lattice while the primary component of the inner shell is a dodecahedral frame composed of dimeric protein P1. For the current study, the 3D architecture of the intact phi 12 virus was obtained by electron cryo-tomography. The nucleocapsid appears to be centered within the membrane envelope and possibly attached to it by bridging structures. Two types of densities were observed protruding from the membrane envelope. The densities of the first type were elongated, running parallel, and closely associated to the envelope outer surface. In contrast, the second density was positioned about 12 nm above the envelope connected to it by a flexible low-density stem. This second structure formed a torroidal structure termed "the donut" and appears to inhibit BHT-induced viral envelope fusion.

Electron cryo-tomographic structure of cystovirus φ12

International Nuclear Information System (INIS)

Hu Guobin; Wei Hui; Rice, William J.; Stokes, David L.; Gottlieb, Paul

2008-01-01

Bacteriophage φ12 is a member of the Cystoviridae virus family and contains a genome consisting of three segments of double-stranded RNA (dsRNA). This virus family contains eight identified members, of which four have been classified in regard to their complete genomic sequence and encoded viral proteins. A phospholipid envelope that contains the integral proteins P6, P9, P10, and P13 surrounds the viral particles. In species φ6, host infection requires binding of a multimeric P3 complex to type IV pili. In species φ8, φ12, and φ13, the attachment apparatus is a heteromeric protein assembly that utilizes the rough lipopolysaccharide (rlps) as a receptor. In φ8 the protein components are designated P3a and P3b while in species φ12 proteins P3a and P3c have been identified in the complex. The phospholipid envelope of the cystoviruses surrounds a nucleocapsid (NC) composed of two shells. The outer shell is composed of protein P8 with a T = 13 icosahedral lattice while the primary component of the inner shell is a dodecahedral frame composed of dimeric protein P1. For the current study, the 3D architecture of the intact φ12 virus was obtained by electron cryo-tomography. The nucleocapsid appears to be centered within the membrane envelope and possibly attached to it by bridging structures. Two types of densities were observed protruding from the membrane envelope. The densities of the first type were elongated, running parallel, and closely associated to the envelope outer surface. In contrast, the second density was positioned about 12 nm above the envelope connected to it by a flexible low-density stem. This second structure formed a torroidal structure termed 'the donut' and appears to inhibit BHT-induced viral envelope fusion

HIE-ISOLDE CRYO-MODULE Assembly - Superconducting Solenoid

CERN Multimedia

Leclercq, Yann

2016-01-01

Assembly of the cryo-module components in SM18 cleanroom. The superconducting solenoid (housed inside its helium vessel) is cleaned, prepared then installed on the supporting frame of the cryo-module and connected to the helium tank, prior to the assembly of the RF cavities on the structure. The completed first 2 cryo-modules installed inside the HIE-ISOLDE-LINAC ready for beam operation is also shown.

CryoSat Plus for Oceans - analysis of the state-of-the-art

Science.gov (United States)

Naeije, Marc; Gommenginger, Christine; Moreau, Thomas; Cotton, David; Benveniste, Jerome; Dinardo Dinardo, Salvatore

2013-04-01

The CryoSat Plus for Oceans (CP4O) project is an ESA initiative carried out by a European wide consortium of altimetry experts. It aims to build a sound scientific basis for new scientific and operational applications of data coming from CryoSat-2 over the open ocean, polar ocean, coastal seas and for seafloor mapping. It also generates and evaluates new methods and products that will enable the full exploitation of the capabilities of the CryoSat-2 SIRAL altimeter, and extend their application beyond the initial mission objectives. It therefore also acts as a preparation for the upcoming Sentinel and Jason SAR enabled altimetry missions. In this paper we address the review of the CryoSat state-of-the-art, relevant current initiatives, algorithms, models and Earth Observation based products and datasets that are relevant in the Cryosat+ ocean theme. Compared to conventional (pulse-limited) altimeter missions, Cryosat-2 is not a dedicated platform for ocean research: typically the microwave radiometer (MWR) for wet tropospheric corrections is lacking, as is the direct measurement of the first order ionospheric effect by means of a dual-frequency altimeter. Also the orbit of Cryosat-2 has a rather long repetition period, unsuited for collinear tracks analyses. These three particular features have been studied already in the HERACLES project on the eve of the first CryoSat launch. We revisit the outcome of this study, update to current understanding and perception, and ultimately develop what was, is and will be proposed in these problem areas. Clearly, we question the standard ionosphere corrections, the wet troposphere corrections and the accuracy of the mean sea surface (MSS) underlying the accuracy of derived sea level anomalies. In addition, Cryosat-2 provides the first innovative altimeter with SAR and SARIn modes. This raises the direct problem of "how to process these data", simply because this has not been done before. Compared to pulse-limited altimetry it

GraDeR: Membrane Protein Complex Preparation for Single-Particle Cryo-EM.

Science.gov (United States)

Hauer, Florian; Gerle, Christoph; Fischer, Niels; Oshima, Atsunori; Shinzawa-Itoh, Kyoko; Shimada, Satoru; Yokoyama, Ken; Fujiyoshi, Yoshinori; Stark, Holger

2015-09-01

We developed a method, named GraDeR, which substantially improves the preparation of membrane protein complexes for structure determination by single-particle cryo-electron microscopy (cryo-EM). In GraDeR, glycerol gradient centrifugation is used for the mild removal of free detergent monomers and micelles from lauryl maltose-neopentyl glycol detergent stabilized membrane complexes, resulting in monodisperse and stable complexes to which standard processes for water-soluble complexes can be applied. We demonstrate the applicability of the method on three different membrane complexes, including the mammalian FoF1 ATP synthase. For this highly dynamic and fragile rotary motor, we show that GraDeR allows visualizing the asymmetry of the F1 domain, which matches the ground state structure of the isolated domain. Therefore, the present cryo-EM structure of FoF1 ATP synthase provides direct structural evidence for Boyer's binding change mechanism in the context of the intact enzyme. Copyright © 2015 Elsevier Ltd. All rights reserved.

Depth sectioning using electron energy loss spectroscopy

International Nuclear Information System (INIS)

D'Alfonso, A J; Findlay, S D; Allen, L J; Cosgriff, E C; Kirkland, A I; Nellist, P D; Oxley, M P

2008-01-01

The continued development of electron probe aberration correctors for scanning transmission electron microscopy has enabled finer electron probes, allowing atomic resolution column-by-column electron energy loss spectroscopy. Finer electron probes have also led to a decrease in the probe depth of focus, facilitating optical slicing or depth sectioning of samples. The inclusion of post specimen aberration corrected image forming lenses allows for scanning confocal electron microscopy with further improved depth resolution and selectivity. We show that in both scanning transmission electron microscopy and scanning confocal electron microscopy geometries, by performing a three dimensional raster scan through a specimen and detecting electrons scattered with a characteristic energy loss, it will be possible to determine the location of isolated impurities embedded within the bulk.

Cryo-electron microscopy of vitrified SV40 minichromosomes: the liquid drop model.

Science.gov (United States)

Dubochet, J; Adrian, M; Schultz, P; Oudet, P

1986-03-01

The structure of SV40 minichromosomes has been studied by cryo-electron microscopy of vitrified thin layers of solution. In high-salt buffer (130 mM NaCl), freshly prepared minichromosomes are condensed into globules 30 nm or more in diameter. On the micrograph, they appear to be formed by the close packing of 10 nm granules which give rise to a 10 nm reflection in the optical diffractogram. The globules can adopt many different conformations. At high concentration, they fuse into a homogeneous 'sea' of closely packed 10 nm granules. In low-salt buffer (less than 10 mM NaCl), the globules open, first into 10 nm filaments, and then into nucleosome-strings. The 'liquid drop' model is proposed to explain the condensed structure of the minichromosome in high-salt buffer: nucleosomes stack specifically on top of one another, thus forming the 10 nm filaments. 10 nm filaments in turn, tend to aggregate laterally. Optimizing both these interactions results in the condensation of 10 nm filaments or portions thereof into a structure similar to that of a liquid. Some implications of this model for the structure of cellular chromatin are discussed.

ESA CryoVEx 2014 - Airborne ASIRAS radar and laser scanner measurements during 2014 CryoVEx campaign in the Arctic

DEFF Research Database (Denmark)

Hvidegaard, S. M.; Nielsen, J. E.; Sørensen, L. Sandberg

the penetration depth of the ASIRAS radar. An opportunity site on the Greenland Ice Sheet was surveyed near Jakobshavn Isbræ. No other ground experiments were coordinated with the CryoVEx campaing on the Greenland Ice Sheet. The CryoVEx 2014 campaign was a success and the processed data is of high quality......This report outlines the airborne field operations with the ESA airborne Ku‐band interferometric radar (ASIRAS), coincident airborne laser scanner (ALS) and vertical photography to acquire data over sea‐ and land ice along validation sites and CryoSat‐2 ground tracks. The airborne campaign...... in the Beaufort Sea lead by US office of Naval Research (ONR) and north of Greenland as a dedicated ESA CryoVEx initiative. In addition, selected CryoSat‐2 ground tracks were under‐flown in the Lincoln Sea from CFS Alert, North of Greenland and Svalbard from St. Nord and Longyearbyen. Several of the flights...

Commissioning result of the KSTAR in-vessel cryo-pump

Energy Technology Data Exchange (ETDEWEB)

Chang, Y. B.; Lee, H. J.; Park, Y.M. [National Fusion Research Institute, Daejeon (Korea, Republic of); and others

2013-12-15

KSTAR in-vessel cryo-pump has been installed in the vacuum vessel top and bottom side with up-down symmetry for the better plasma density control in the D-shape H-mode. The cryogenic helium lines of the in-vessel cryo-pump are located at the vertical positions from the vacuum vessel torus center 2,000 mm. The inductive electrical potential has been optimized to reduce risk of electrical breakdown during plasma disruption. In-vessel cryo-pump consists of three parts of coaxial circular shape components; cryo-panel, thermal shield and particle shield. The cryo-panel is cooled down to below 4.5 K. The cryo-panel and thermal shields were made by Inconel 625 tube for higher mechanical strength. The thermal shields and their cooling tubes were annealed in air environment to improve the thermal radiation emissivity on the surface. Surface of cryo-panel was electro-polished to minimize the thermal radiation heat load. The in-vessel cryo-pump was pre-assembled on a test bed in 180 degree segment base. The leak test was carried out after the thermal shock between room temperature to LN2 one before installing them into vacuum vessel. Two segments were welded together in the vacuum vessel and final leak test was performed after the thermal shock. Commissioning of the in-vessel cryo-pump was carried out using a temporary liquid helium supply system.

Molecular dynamics-based refinement and validation for sub-5 Å cryo-electron microscopy maps.

Science.gov (United States)

Singharoy, Abhishek; Teo, Ivan; McGreevy, Ryan; Stone, John E; Zhao, Jianhua; Schulten, Klaus

2016-07-07

Two structure determination methods, based on the molecular dynamics flexible fitting (MDFF) paradigm, are presented that resolve sub-5 Å cryo-electron microscopy (EM) maps with either single structures or ensembles of such structures. The methods, denoted cascade MDFF and resolution exchange MDFF, sequentially re-refine a search model against a series of maps of progressively higher resolutions, which ends with the original experimental resolution. Application of sequential re-refinement enables MDFF to achieve a radius of convergence of ~25 Å demonstrated with the accurate modeling of β-galactosidase and TRPV1 proteins at 3.2 Å and 3.4 Å resolution, respectively. The MDFF refinements uniquely offer map-model validation and B-factor determination criteria based on the inherent dynamics of the macromolecules studied, captured by means of local root mean square fluctuations. The MDFF tools described are available to researchers through an easy-to-use and cost-effective cloud computing resource on Amazon Web Services.

Attainment of 40.5 pm spatial resolution using 300 kV scanning transmission electron microscope equipped with fifth-order aberration corrector.

Science.gov (United States)

Morishita, Shigeyuki; Ishikawa, Ryo; Kohno, Yuji; Sawada, Hidetaka; Shibata, Naoya; Ikuhara, Yuichi

2018-02-01

The achievement of a fine electron probe for high-resolution imaging in scanning transmission electron microscopy requires technological developments, especially in electron optics. For this purpose, we developed a microscope with a fifth-order aberration corrector that operates at 300 kV. The contrast flat region in an experimental Ronchigram, which indicates the aberration-free angle, was expanded to 70 mrad. By using a probe with convergence angle of 40 mrad in the scanning transmission electron microscope at 300 kV, we attained the spatial resolution of 40.5 pm, which is the projected interatomic distance between Ga-Ga atomic columns of GaN observed along [212] direction.

CryoSat product quality and evolutions

Science.gov (United States)

Bouffard, Jérome; Parrinello, Tommaso; Féménias, Pierre

2017-04-01

The main CryoSat mission objectives are to measure the regional and basin-scale changes in the thickness of the sea-ice and, in the elevation of the ice sheets and mountain glaciers. Beside its ice-monitoring objective, CryoSat also provides valuable observations for the oceanographic community. The CryoSat data are processed by ESA both over the ocean and ice surfaces with two independent processors. These data need to be routinely Quality-Controlled and thoroughly Validated (QCV). Based on the QCV outcomes from ESA multi-national partners and the feedback from the scientific community, the data products continuously evolve in order to accommodate a wide range of users over the Sea ice, the Land Ice and the Ocean domains. The main objectives of this paper are to give an overview of main CryoSat QCV results and product improvements; as well as to present the processing algorithm upgrades being implemented for future ice and ocean Baselines.

CryoCart Restoration and Vacuum Pipe Construction

Science.gov (United States)

Chaidez, Mariana

2016-01-01

first completed at the component level. During this process, the igniter of the main engine and the RCS thrusters will be tested under a vacuum. To complete the testing of the components, the test setup first needed to be finalized. The CryoCart is being used to feed the propellants to the test article. The CryoCart is a movable test set-up that was developed in 2009 to provide a mobile platform for testing oxygen/methane systems with hot-fire capability up to 100 lbf. The CryoCart consists of three different systems: Oxygen, Methane, and liquid Nitrogen. The Oxygen and Methane systems are placed into two different carts while the liquid nitrogen system is mainly located in the methane cart. Over the years, the CryoCart has been utilized for different projects and has undergone deterioration. For this reason, a new phase has been developed to rebuild it to working conditions once again. During my internship, I was aiding in the construction and restoration of the CryoCart. In the initial stages of the process, I updated the fluid and electrical schematics for the oxygen, methane, and test article systems. The original CryoCart consisted of an electrical panel that utilized electromechanical relays and a terminal to drive the igniter power and signal, as well as the main fuel and oxygen valves. This electrical panel connected to the CryoCart through various wire harnesses that could be found exiting from the CryoCart. First, it was determined how these harnesses connected to the electromechanical relays so that they worked correctly. Once the electrical system was understood, an alternative for the electromechanical relays and the Molex connectors used throughout the system was sought since these components can often prove to be unreliable. Solid State relays and MIL connectors were purchased to serve as replacements. Upon arrival of the parts, crimping and wiring was completed to install the new solid state relays and MIL connectors. During the replacement of the relays

Comparison of CryoSat-2 and ENVISAT radar freeboard over Arctic sea ice: toward an improved Envisat freeboard retrieval

Science.gov (United States)

Guerreiro, Kevin; Fleury, Sara; Zakharova, Elena; Kouraev, Alexei; Rémy, Frédérique; Maisongrande, Philippe

2017-09-01

Over the past decade, sea-ice freeboard has been monitored with various satellite altimetric missions with the aim of producing long-term time series of ice thickness. While recent studies have demonstrated the capacity of the CryoSat-2 mission (2010-present) to provide accurate freeboard measurements, the current estimates obtained with the Envisat mission (2002-2012) still require some large improvements. In this study, we first estimate Envisat and CryoSat-2 radar freeboard by using the exact same processing algorithms. We then analyse the freeboard difference between the two estimates over the common winter periods (November 2010-April 2011 and November 2011-March 2012). The analysis of along-track data and gridded radar freeboard in conjunction with Envisat pulse-peakiness (PP) maps suggests that the discrepancy between the two sensors is related to the surface properties of sea-ice floes and to the use of a threshold retracker. Based on the relation between the Envisat pulse peakiness and the radar freeboard difference between Envisat and CryoSat-2, we produce a monthly CryoSat-2-like version of Envisat freeboard. The improved Envisat data set freeboard displays a similar spatial distribution to CryoSat-2 (RMSD = 1.5 cm) during the two ice growth seasons and for all months of the period of study. The comparison of the altimetric data sets with in situ ice draught measurements during the common flight period shows that the improved Envisat data set (RMSE = 12-28 cm) is as accurate as CryoSat-2 (RMSE = 15-21 cm) and much more accurate than the uncorrected Envisat data set (RMSE = 178-179 cm). The comparison of the improved Envisat radar freeboard data set is then extended to the rest of the Envisat mission to demonstrate the validity of PP correction from the calibration period. The good agreement between the improved Envisat data set and the in situ ice draught data set (RMSE = 13-32 cm) demonstrates the potential of the PP correction to produce accurate

Brillouin micro-spectroscopy through aberrations via sensorless adaptive optics

Science.gov (United States)

Edrei, Eitan; Scarcelli, Giuliano

2018-04-01

Brillouin spectroscopy is a powerful optical technique for non-contact viscoelastic characterizations which has recently found applications in three-dimensional mapping of biological samples. Brillouin spectroscopy performances are rapidly degraded by optical aberrations and have therefore been limited to homogenous transparent samples. In this work, we developed an adaptive optics (AO) configuration designed for Brillouin scattering spectroscopy to engineer the incident wavefront and correct for aberrations. Our configuration does not require direct wavefront sensing and the injection of a "guide-star"; hence, it can be implemented without the need for sample pre-treatment. We used our AO-Brillouin spectrometer in aberrated phantoms and biological samples and obtained improved precision and resolution of Brillouin spectral analysis; we demonstrated 2.5-fold enhancement in Brillouin signal strength and 1.4-fold improvement in axial resolution because of the correction of optical aberrations.

Aberration compensation using a spatial light modulator LCD

International Nuclear Information System (INIS)

Amezquita, R; Rincon, O; Torres, Y M

2011-01-01

The dynamic correction of aberrations introduced in optical systems have been a widely discussed topic in the past 10 years. Adaptive optics is the most important developed field where the Shack-Hartmann sensors and deformable mirrors are used for the measurement and correction of wavefronts. In this paper, an interferometric set-up which uses a Spatial Light Modulator (SLM) as an active element is proposed. Using this SLM a procedure for the compensation of all phase aberrations present in the experimental setup is shown.

An adjustable electron achromat for cathode lens microscopy

Energy Technology Data Exchange (ETDEWEB)

Tromp, R.M., E-mail: rtromp@us.ibm.com [IBM T.J. Watson Research Center, 1101 Kitchawan Road, Yorktown Heights, NY 10598 (United States); Leiden Institute of Physics, Kamerlingh Onnes Laboratory, Niels Bohrweg 2, 2333 CA Leiden (Netherlands)

2015-12-15

Chromatic aberration correction in light optics began with the invention of a two-color-corrected achromatic crown/flint lens doublet by Chester Moore Hall in 1730. Such color correction is necessary because any single glass shows dispersion (i.e. its index of refraction changes with wavelength), which can be counteracted by combining different glasses with different dispersions. In cathode lens microscopes (such as Photo Electron Emission Microscopy – PEEM) we encounter a similar situation, where the chromatic aberration coefficient of the cathode lens shows strong dispersion, i.e. depends (non-linearly) on the energy with which the electrons leave the sample. Here I show how a cathode lens in combination with an electron mirror can be configured as an adjustable electron achromat. The lens/mirror combination can be corrected at two electron energies by balancing the settings of the electron mirror against the settings of the cathode lens. The achromat can be adjusted to deliver optimum performance, depending on the requirements of a specific experiment. Going beyond the achromat, an apochromat would improve resolution and transmission by a very significant margin. I discuss the requirements and outlook for such a system, which for now remains a wish waiting for fulfilment. - Highlights: • The properties of cathode objective lens plus electron mirror are discussed. • In analogy with light-optical achromats, cathode lens plus mirror can be configured as an electron achromat. • Unlike light optics, the electron achromat can be adjusted to best fulfill experimental requirements.

Atomic Resolution Structures of Human Bufaviruses Determined by Cryo-Electron Microscopy

Directory of Open Access Journals (Sweden)

Maria Ilyas

2018-01-01

Full Text Available Bufavirus strain 1 (BuV1, a member of the Protoparvovirus genus of the Parvoviridae, was first isolated from fecal samples of children with acute diarrhea in Burkina Faso. Since this initial discovery, BuVs have been isolated in several countries, including Finland, the Netherlands, and Bhutan, in pediatric patients exhibiting similar symptoms. Towards their characterization, the structures of virus-like particles of BuV1, BuV2, and BuV3, the current known genotypes, have been determined by cryo-electron microscopy and image reconstruction to 2.84, 3.79, and 3.25 Å, respectively. The BuVs, 65–73% identical in amino acid sequence, conserve the major viral protein, VP2, structure and general capsid surface features of parvoviruses. These include a core β-barrel (βB-βI, α-helix A, and large surface loops inserted between these elements in VP2. The capsid contains depressions at the icosahedral 2-fold and around the 5-fold axes, and has three separated protrusions surrounding the 3-fold axes. Structure comparison among the BuVs and to available parvovirus structures revealed capsid surface variations and capsid 3-fold protrusions that depart from the single pinwheel arrangement of the animal protoparvoviruses. These structures provide a platform to begin the molecular characterization of these potentially pathogenic viruses.

New design for a space cryo-mechanism

Science.gov (United States)

Durand, Gilles; Amiaux, Jérôme; Augueres, Jean-Louis; Carty, Michael; Barrière, Jean Christophe; Bouzat, Marylène; Duboué, Bruno; Lagage, Pierre Olivier; Lebeuf, Didier; Lepage, Erwan; Lemer, Isabelle; Marlaguey, Nathalie Peydrol; Poupar, Sébastien

2008-07-01

Based on its experience of space application instrument and its development of cryomechanism for astronomical ground based instrument VLT / VISIR, CEA Saclay is proposing a new concept of Space Cryomechanism. This design is based on VLT/VISIR cryo-mechanism design adapted to space requirements taking into account all the specification of space environment (vibrations at launch, cryogenic vacuum, materials, radiations, ...). The original concept of the design is based on the association of the key elements: a dog-clutch with Hirth teeth jaws coupled to a step-by-step space qualified cryo-motor, a bellows that allows for separation of indexing and rotating functions, and enlarged bearings design in "O" arrangement that increase robustness to vibration. The actuator has 360 steady positions that can be reached within les than a second with repeatability of 5 arcsec peak to peak. After a presentation of the details of the concept and of its benefits to robustness to space environment, the paper describes the thoroughly qualification program of the cryo-mechanism with respect to space requirements (cryo-cycling, indexing accuracy, power consumption, heat dissipation, motorisation margins, vibrations). This cryo-mechanism may be built in 3 different sizes for wheels up to 10 kg.

Cryo-balloon catheter localization in fluoroscopic images

Science.gov (United States)

Kurzendorfer, Tanja; Brost, Alexander; Jakob, Carolin; Mewes, Philip W.; Bourier, Felix; Koch, Martin; Kurzidim, Klaus; Hornegger, Joachim; Strobel, Norbert

2013-03-01

Minimally invasive catheter ablation has become the preferred treatment option for atrial fibrillation. Although the standard ablation procedure involves ablation points set by radio-frequency catheters, cryo-balloon catheters have even been reported to be more advantageous in certain cases. As electro-anatomical mapping systems do not support cryo-balloon ablation procedures, X-ray guidance is needed. However, current methods to provide support for cryo-balloon catheters in fluoroscopically guided ablation procedures rely heavily on manual user interaction. To improve this, we propose a first method for automatic cryo-balloon catheter localization in fluoroscopic images based on a blob detection algorithm. Our method is evaluated on 24 clinical images from 17 patients. The method successfully detected the cryoballoon in 22 out of 24 images, yielding a success rate of 91.6 %. The successful localization achieved an accuracy of 1.00 mm +/- 0.44 mm. Even though our methods currently fails in 8.4 % of the images available, it still offers a significant improvement over manual methods. Furthermore, detecting a landmark point along the cryo-balloon catheter can be a very important step for additional post-processing operations.

CryoSat-2: A new perspective on the Cryosphere

Science.gov (United States)

Shepherd, A.; Armitage, T.; Briggs, K.; Flament, T.; Hogg, A. E.; McMillan, M.; Muir, A.; Ridout, A.; Sundal, A.; Tilling, R.; Wingham, D.

2014-12-01

CryoSat-2 is ESA's first satellite mission dedicated to measuring changes in the polar land ice and sea ice cover. Following its launch in April 2010, we have examined the performance of the instrument over the continental ice sheets of Antarctica and Greenland, the Artic Ocean, and, for the purposes of calibration, over the oceans. We have confirmed the engineering performance at system level of the interferometer demonstrating that it measures across-track surface slopes with a precision of 25 micro-radians and an accuracy of 10 micro-radians, greatly exceeding the pre-launch specifications (100 micro-radians). Over the polar ice sheets, we have examined the performance of the range estimation, and determined the range precision to be 19 cm RMS at 20 Hz. We have examined the retrieval of the phase information over the ice sheets, and found the phase estimates to be robust and little affected by the uncertain ice sheet topography. Based on the calibration of the interferometer, the contribution of the across track slope error is, at 0.4 mm, negligible. Over marine sea ice, we have verified the discrimination of sea ice and ocean lead returns using contemporaneous SAR imagery from ENVISAT, and we have estimated the precision of individual (20 Hz) measurements to be 2 cm. In summary, with the corrected data products, we are able to confirm that the system performance of CryoSat-2 will meet or exceed its specification over the continental and marine ice sheets. This presenetation sumamrises the key outcomes of the mission performance, and presents a series of example case studies where CryoSat-2 data have been applied to study changes in Earth's land and sea ice cover. We show that in 4 years CryoSat has been able to detect changes in the mass of the Antarctic and Greenland ice sheets with an accuracy comparable to that of the past 20 years of conventional satellite altimetry and that important changes have occured in these regions, we show that CryoSat has been able

Cryo-scanning electron microscopy discloses differences in dehydration of frozen boar semen stored in large containers.

Science.gov (United States)

Ekwall, H

2009-02-01

In general, freezing in flat plastic polyethylene terephthalate (PET) bags (FlatPacks) at 50 degrees C/min gives better post-thaw viability, in terms of sperm motility and membrane integrity, than does freezing in plastic maxi-straws, probably owing to differences in cryobiology. To test the hypothesis that this better survival post-thaw relates to the degree of sperm dehydration during freezing, the present study investigated the structure of boar semen in a frozen state using cryo-scanning electron microscopy (cryo-SEM) to compare two different packages (FlatPacks and maxi-straws) for single artificial insemination (AI) doses, and three different freezing rates. The semen was split-sample frozen in maxi-straws or FlatPacks (both holding 5 ml) using 3% glycerol as cryoprotectant. Three freezing rates were applied from -5 degrees C to -100 degrees C, namely 2 degrees C/min, 50 degrees C/min and 1200 degrees C/min, the lattermost by plunging the samples into liquid nitrogen (LN(2)). The samples were thereafter fractured into LN(2) and larger areas of extra-cellular, unbound frozen water ('ice lakes') were measured to determine the degree of dehydration of the spermatozoa. These areas decreased in size with an increase in cooling rate, the differences in size being more dramatic for maxi-straws than for FlatPacks. Size of ice lakes was also influenced by location within package in relation to cooling rate, the central values being always smaller in maxi-straws than in Flatpacks (p < 0.05 at 2 degrees C/min and 50 degrees C/min) but not at 1200 degrees C/min, which suggested the FlatPack allows for more homogenous freezing of boar semen.

Thermo-mechanical design methodology for ITER cryo-distribution cold boxes

International Nuclear Information System (INIS)

Shukla, Vinit; Patel, Pratik; Vaghela, Hiten; Das, Jotirmoy; Shah, Nitin; Bhattacharya, Ritendra; Sarkar, Biswanath; Chang, Hyun-sik

2015-01-01

The ITER cryo-distribution system is in charge of the proper distribution of the cryogen at required mass flow rate, pressure and temperature level to the users namely; the superconducting magnets and cryopumps. The cryo-distribution also acts as a thermal buffer in order to run the cryo-plant as much as possible at a steady state condition. A typical cryo-distribution cold box is equipped with mainly liquid helium bath with heat exchangers, cryogenic valves, cold circulating pump and cold compressor. During the intended operation life of ITER, several loads on the cryo-distribution system are envisaged, these are, gravity/assembly loads, nominal pressure/temperature, test pressure/temperature, purge pressure, thermo-mechanical loads due to break of insulation vacuum, transport acceleration and seismic loads. Single loads or combinations of them can act on the cryo-distribution system and its components; therefore, it is very important to analyze the behavior of the system and components under the influence of these loads or combinations. Possible load combinations for the cryo-distribution system will be analyzed and will lead to the basis of the design. This paper will focus on the understanding of the nature of the loads and their combinations for the ITER cryo-distribution as well as their impacts on the design. A representative model of a cold box is considered on which the load combinations have been applied in order to understand their impacts on the design of the cryo-distribution. Also the worst-impact loads or their combination which drive the design of cryo-distribution cold boxes will be derived. (author)

The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm.

Science.gov (United States)

Hrubanova, Kamila; Nebesarova, Jana; Ruzicka, Filip; Krzyzanek, Vladislav

2018-04-22

In this study we present an innovative method for the preparation of fully hydrated samples of microbial biofilms of cultures Staphylococcus epidermidis, Candida parapsilosis and Candida albicans. Cryo-scanning electron microscopy (cryo-SEM) and high-pressure freezing (HPF) rank among cutting edge techniques in the electron microscopy of hydrated samples such as biofilms. However, the combination of these techniques is not always easily applicable. Therefore, we present a method of combining high-pressure freezing using EM PACT2 (Leica Microsystems), which fixes hydrated samples on small sapphire discs, with a high resolution SEM equipped with the widely used cryo-preparation system ALTO 2500 (Gatan). Using a holder developed in house, a freeze-fracturing technique was applied to image and investigate microbial cultures cultivated on the sapphire discs. In our experiments, we focused on the ultrastructure of the extracellular matrix produced during cultivation and the relationships among microbial cells in the biofilm. The main goal of our investigations was the detailed visualization of areas of the biofilm where the microbial cells adhere to the substrate/surface. We show the feasibility of this technique, which is clearly demonstrated in experiments with various freeze-etching times. Copyright © 2018 Elsevier Ltd. All rights reserved.

Cryo-immunogold electron microscopy for prions: toward identification of a conversion site.

Science.gov (United States)

Godsave, Susan F; Wille, Holger; Kujala, Pekka; Latawiec, Diane; DeArmond, Stephen J; Serban, Ana; Prusiner, Stanley B; Peters, Peter J

2008-11-19

Prion diseases are caused by accumulation of an abnormally folded isoform (PrP(Sc)) of the cellular prion protein (PrP(C)). The subcellular distribution of PrP(Sc) and the site of its formation in brain are still unclear. We performed quantitative cryo-immunogold electron microscopy on hippocampal sections from mice infected with the Rocky Mountain Laboratory strain of prions. Two antibodies were used: R2, which recognizes both PrP(C) and PrP(Sc); and F4-31, which only detects PrP(C) in undenatured sections. At a late subclinical stage of prion infection, both PrP(C) and PrP(Sc) were detected principally on neuronal plasma membranes and on vesicles resembling early endocytic or recycling vesicles in the neuropil. The R2 labeling was approximately six times higher in the infected than the uninfected hippocampus and gold clusters were only evident in infected tissue. The biggest increase in labeling density (24-fold) was found on the early/recycling endosome-like vesicles of small-diameter neurites, suggesting these as possible sites of conversion. Trypsin digestion of infected hippocampal sections resulted in a reduction in R2 labeling of >85%, which suggests that a high proportion of PrP(Sc) may be oligomeric, protease-sensitive PrP(Sc).

Cryo-EM structures of two bovine adenovirus type 3 intermediates

International Nuclear Information System (INIS)

Cheng, Lingpeng; Huang, Xiaoxing; Li, Xiaomin; Xiong, Wei; Sun, Wei; Yang, Chongwen; Zhang, Kai; Wang, Ying; Liu, Hongrong; Huang, Xiaojun; Ji, Gang; Sun, Fei; Zheng, Congyi; Zhu, Ping

2014-01-01

Adenoviruses (Ads) infect hosts from all vertebrate species and have been investigated as vaccine vectors. We report here near-atomic structures of two bovine Ad type 3 (BAd3) intermediates obtained by cryo-electron microscopy. A comparison between the two intermediate structures reveals that the differences are localized in the fivefold vertex region, while their facet structures are identical. The overall facet structure of BAd3 exhibits a similar structure to human Ads; however, BAd3 protein IX has a unique conformation. Mass spectrometry and cryo-electron tomography analyses indicate that one intermediate structure represents the stage during DNA encapsidation, whilst the other intermediate structure represents a later stage. These results also suggest that cleavage of precursor protein VI occurs during, rather than after, the DNA encapsidation process. Overall, our results provide insights into the mechanism of Ad assembly, and allow the first structural comparison between human and nonhuman Ads at backbone level. - Highlights: • First structure of bovine adenovirus type 3. • Some channels are located at the vertex of intermediate during DNA encapsidation. • Protein IX exhibits a unique conformation of trimeric coiled–coiled structure. • Cleavage of precursor protein VI occurs during the DNA encapsidation process

Cryo-EM structures of two bovine adenovirus type 3 intermediates

Energy Technology Data Exchange (ETDEWEB)

Cheng, Lingpeng; Huang, Xiaoxing; Li, Xiaomin [National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); Xiong, Wei [State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Luo-jia-shan, Wuhan, Hubei 430072 (China); Sun, Wei; Yang, Chongwen; Zhang, Kai; Wang, Ying [National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); Liu, Hongrong [College of Physics and Information Science, Hunan Normal University, Changsha, Hunan 410081 (China); Huang, Xiaojun; Ji, Gang; Sun, Fei [National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); Zheng, Congyi, E-mail: cctcc202@whu.edu.cn [State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Luo-jia-shan, Wuhan, Hubei 430072 (China); Zhu, Ping, E-mail: zhup@ibp.ac.cn [National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China)

2014-02-15

Adenoviruses (Ads) infect hosts from all vertebrate species and have been investigated as vaccine vectors. We report here near-atomic structures of two bovine Ad type 3 (BAd3) intermediates obtained by cryo-electron microscopy. A comparison between the two intermediate structures reveals that the differences are localized in the fivefold vertex region, while their facet structures are identical. The overall facet structure of BAd3 exhibits a similar structure to human Ads; however, BAd3 protein IX has a unique conformation. Mass spectrometry and cryo-electron tomography analyses indicate that one intermediate structure represents the stage during DNA encapsidation, whilst the other intermediate structure represents a later stage. These results also suggest that cleavage of precursor protein VI occurs during, rather than after, the DNA encapsidation process. Overall, our results provide insights into the mechanism of Ad assembly, and allow the first structural comparison between human and nonhuman Ads at backbone level. - Highlights: • First structure of bovine adenovirus type 3. • Some channels are located at the vertex of intermediate during DNA encapsidation. • Protein IX exhibits a unique conformation of trimeric coiled–coiled structure. • Cleavage of precursor protein VI occurs during the DNA encapsidation process.

Double deflection system for an electron beam device

International Nuclear Information System (INIS)

Parker, N.W.; Crewe, A.V.

1978-01-01

A double deflection scanning system for electron beam instruments is provided embodying a means of correcting isotropic coma, and anisotropic coma aberrations induced by the magnetic lens of such an instrument. The scanning system deflects the beam prior to entry into the magnetic lens from the normal on-axis intersection of the beam with the lens according to predetermined formulas and thereby reduces the aberrations

Development of forage cryo minced fish technology

Directory of Open Access Journals (Sweden)

Titova S. A.

2017-11-01

Full Text Available The technology of grinding the frozen fish raw material using the useful model of a piston-type extrusion plant with cold working bodies for fodder minced fish (cryo minced fish feed by the method of cryoextrusion has been developed. The possibility of replacing the standard manufacturing operations (defrosting and subsequent grinding of raw materials to the one (cryoextrusion has been substantiated. The optimal shape of holes of the die for forcing the raw material has been determined in the form of "hourglass", the hole diameter is 7 mm, the length of the process of pushing is 40 s. It has been established that the finished product (cryo minced fish feed has a homogeneous grinding, juicy, crumbly texture, the temperature in the thickness of the product is equal to the temperature of the raw material before processing (–18 °C. The technological scheme of producing the minced fish feed by the method of cryoextrusion has been developed. Researching the chemical composition of fish and feed of mince produced by the traditional and developed methods it has been established that protein content in the cryo minced fish feed remains unchanged. The amino acid composition of cryo minced fish feed has been determined, the amount of essential amino acids is 365.6 mg/g of protein. In the proteins of the cryo minced fish feed methionine has been contained in the minimum quantity (10.0 mg/g protein and in the maximum – serine (155.3 mg/g protein. In the protein of the cryo minced fish feed the only limiting amino acid is methionine. The coefficient of rationality of the cryo minced fish feed whiting is equal to 0.33. The product is not toxic, is not subject to contamination with dangerous microorganisms, has high biological value, contains necessary in animal nutrition mineral elements, the energy value per 100 g of the product is 87 kcal. The combination of the processes of cryoextrusion and lyophilization allows to obtain a new food product with high

Airborne campaigns for CryoSat pre-launch calibration and validation

DEFF Research Database (Denmark)

Hvidegaard, Sine Munk; Forsberg, René; Skourup, Henriette

2010-01-01

From 2003 to 2008 DTU Space together with ESA and several international partners carried out airborne and ground field campaigns in preparation for CryoSat validation; called CryoVEx: CryoSat Validation Experiments covering the main ice caps in Greenland, Canada and Svalbard and sea ice in the Ar......From 2003 to 2008 DTU Space together with ESA and several international partners carried out airborne and ground field campaigns in preparation for CryoSat validation; called CryoVEx: CryoSat Validation Experiments covering the main ice caps in Greenland, Canada and Svalbard and sea ice...... in the Arctic Ocean. The main goal of the airborne surveys was to acquire coincident scanning laser and CryoSat type radar elevation measurements of the surface; either sea ice or land ice. Selected lines have been surveyed along with detailed mapping of validation sites coordinated with insitu field work...... and helicopter electromagnetic surveying. This paper summarises the pre-launch campaigns and presents some of the result from the coincident measurement from airborne and ground observations....

A versatile atomic number correction for electron-probe microanalysis

International Nuclear Information System (INIS)

Love, G.; Cox, M.G.; Scott, V.D.

1978-01-01

A new atomic number correction is proposed for quantitative electron-probe microanalysis. Analytical expressions for the stopping power S and back-scatter R factors are derived which take into account atomic number of the target, incident electron energy and overvoltage; the latter expression is established using Monte Carlo calculations. The correct procedures for evaluating S and R for multi-element specimens are described. The new method, which overcomes some limitations inherent in earlier atomic number corrections, may readily be used where specimens are inclined to the electron beam. (author)

Electron holography for fields in solids: problems and progress.

Science.gov (United States)

Lichte, Hannes; Börrnert, Felix; Lenk, Andreas; Lubk, Axel; Röder, Falk; Sickmann, Jan; Sturm, Sebastian; Vogel, Karin; Wolf, Daniel

2013-11-01

Electron holography initially was invented by Dennis Gabor for solving the problems raised by the aberrations of electron lenses in Transmission Electron Microscopy. Nowadays, after hardware correction of aberrations allows true atomic resolution of the structure, for comprehensive understanding of solids, determination of electric and magnetic nanofields is the most challenging task. Since fields are phase objects in the TEM, electron holography is the unrivaled method of choice. After more than 40 years of experimental realization and steady improvement, holography is increasingly contributing to these highly sophisticated and essential questions in materials science, as well to the understanding of electron waves and their interaction with matter. © 2013 Elsevier B.V. All rights reserved.

Estimation of dose from chromosome aberration rate

International Nuclear Information System (INIS)

Li Deping

1990-01-01

The methods and skills of evaluating dose from correctly scored shromsome aberration rate are presented, and supplemented with corresponding BASIC computer code. The possibility and preventive measures of excessive probability of missing score of the aberrations in some of the current routine score methods are discussed. The use of dose-effect relationship with exposure time correction factor G in evaluating doses and their confidence intervals, dose estimation in mixed n-γ exposure, and identification of high by nonuniform acute exposure to low LET radiation and its dose estimation are discussed in more detail. The difference of estimated dose due to whether the interaction between subleisoms produced by n and γ have been taken into account is examined. In fitting the standard dose-aberration rate curve, proper weighing of experiment points and comparison with commonly accepted values are emphasised, and the coefficient of variation σ y √y of the aberration rate y as a function of dose and exposure time is given. In appendix I and II, the dose-aberration rate formula is derived from dual action theory, and the time variation of subleisom is illustrated and in appendix III, the estimation of dose from scores of two different types of aberrations (of other related score) is illustrated. Two computer codes are given in appendix IV, one is a simple code, the other a complete code, including the fitting of standard curve. the skills of using compressed data storage, and the production of simulated 'data ' for testing the curve fitting procedure are also given

Big data in cryoEM: automated collection, processing and accessibility of EM data.

Science.gov (United States)

Baldwin, Philip R; Tan, Yong Zi; Eng, Edward T; Rice, William J; Noble, Alex J; Negro, Carl J; Cianfrocco, Michael A; Potter, Clinton S; Carragher, Bridget

2018-06-01

The scope and complexity of cryogenic electron microscopy (cryoEM) data has greatly increased, and will continue to do so, due to recent and ongoing technical breakthroughs that have led to much improved resolutions for macromolecular structures solved using this method. This big data explosion includes single particle data as well as tomographic tilt series, both generally acquired as direct detector movies of ∼10-100 frames per image or per tilt-series. We provide a brief survey of the developments leading to the current status, and describe existing cryoEM pipelines, with an emphasis on the scope of data acquisition, methods for automation, and use of cloud storage and computing. Copyright © 2017 Elsevier Ltd. All rights reserved.

Transcranial passive acoustic mapping with hemispherical sparse arrays using CT-based skull-specific aberration corrections: a simulation study

International Nuclear Information System (INIS)

Jones, Ryan M; O’Reilly, Meaghan A; Hynynen, Kullervo

2013-01-01

The feasibility of transcranial passive acoustic mapping with hemispherical sparse arrays (30 cm diameter, 16 to 1372 elements, 2.48 mm receiver diameter) using CT-based aberration corrections was investigated via numerical simulations. A multi-layered ray acoustic transcranial ultrasound propagation model based on CT-derived skull morphology was developed. By incorporating skull-specific aberration corrections into a conventional passive beamforming algorithm (Norton and Won 2000 IEEE Trans. Geosci. Remote Sens. 38 1337–43), simulated acoustic source fields representing the emissions from acoustically-stimulated microbubbles were spatially mapped through three digitized human skulls, with the transskull reconstructions closely matching the water-path control images. Image quality was quantified based on main lobe beamwidths, peak sidelobe ratio, and image signal-to-noise ratio. The effects on the resulting image quality of the source’s emission frequency and location within the skull cavity, the array sparsity and element configuration, the receiver element sensitivity, and the specific skull morphology were all investigated. The system’s resolution capabilities were also estimated for various degrees of array sparsity. Passive imaging of acoustic sources through an intact skull was shown possible with sparse hemispherical imaging arrays. This technique may be useful for the monitoring and control of transcranial focused ultrasound (FUS) treatments, particularly non-thermal, cavitation-mediated applications such as FUS-induced blood–brain barrier disruption or sonothrombolysis, for which no real-time monitoring techniques currently exist. (paper)

Transcranial passive acoustic mapping with hemispherical sparse arrays using CT-based skull-specific aberration corrections: a simulation study

Science.gov (United States)

Jones, Ryan M.; O’Reilly, Meaghan A.; Hynynen, Kullervo

2013-01-01

The feasibility of transcranial passive acoustic mapping with hemispherical sparse arrays (30 cm diameter, 16 to 1372 elements, 2.48 mm receiver diameter) using CT-based aberration corrections was investigated via numerical simulations. A multi-layered ray acoustic transcranial ultrasound propagation model based on CT-derived skull morphology was developed. By incorporating skull-specific aberration corrections into a conventional passive beamforming algorithm (Norton and Won 2000 IEEE Trans. Geosci. Remote Sens. 38 1337–43), simulated acoustic source fields representing the emissions from acoustically-stimulated microbubbles were spatially mapped through three digitized human skulls, with the transskull reconstructions closely matching the water-path control images. Image quality was quantified based on main lobe beamwidths, peak sidelobe ratio, and image signal-to-noise ratio. The effects on the resulting image quality of the source’s emission frequency and location within the skull cavity, the array sparsity and element configuration, the receiver element sensitivity, and the specific skull morphology were all investigated. The system’s resolution capabilities were also estimated for various degrees of array sparsity. Passive imaging of acoustic sources through an intact skull was shown possible with sparse hemispherical imaging arrays. This technique may be useful for the monitoring and control of transcranial focused ultrasound (FUS) treatments, particularly non-thermal, cavitation-mediated applications such as FUS-induced blood-brain barrier disruption or sonothrombolysis, for which no real-time monitoring technique currently exists. PMID:23807573

Adaptive compensation of aberrations in ultrafast 3D microscopy using a deformable mirror

Science.gov (United States)

Sherman, Leah R.; Albert, O.; Schmidt, Christoph F.; Vdovin, Gleb V.; Mourou, Gerard A.; Norris, Theodore B.

2000-05-01

3D imaging using a multiphoton scanning confocal microscope is ultimately limited by aberrations of the system. We describe a system to adaptively compensate the aberrations with a deformable mirror. We have increased the transverse scanning range of the microscope by three with compensation of off-axis aberrations.We have also significantly increased the longitudinal scanning depth with compensation of spherical aberrations from the penetration into the sample. Our correction is based on a genetic algorithm that uses second harmonic or two-photon fluorescence signal excited by femtosecond pulses from the sample as the enhancement parameter. This allows us to globally optimize the wavefront without a wavefront measurement. To improve the speed of the optimization we use Zernike polynomials as the basis for correction. Corrections can be stored in a database for look-up with future samples.

Comparison of CryoSat-2 and ENVISAT radar freeboard over Arctic sea ice: toward an improved Envisat freeboard retrieval

Directory of Open Access Journals (Sweden)

K. Guerreiro

2017-09-01

Full Text Available Over the past decade, sea-ice freeboard has been monitored with various satellite altimetric missions with the aim of producing long-term time series of ice thickness. While recent studies have demonstrated the capacity of the CryoSat-2 mission (2010–present to provide accurate freeboard measurements, the current estimates obtained with the Envisat mission (2002–2012 still require some large improvements. In this study, we first estimate Envisat and CryoSat-2 radar freeboard by using the exact same processing algorithms. We then analyse the freeboard difference between the two estimates over the common winter periods (November 2010–April 2011 and November 2011–March 2012. The analysis of along-track data and gridded radar freeboard in conjunction with Envisat pulse-peakiness (PP maps suggests that the discrepancy between the two sensors is related to the surface properties of sea-ice floes and to the use of a threshold retracker. Based on the relation between the Envisat pulse peakiness and the radar freeboard difference between Envisat and CryoSat-2, we produce a monthly CryoSat-2-like version of Envisat freeboard. The improved Envisat data set freeboard displays a similar spatial distribution to CryoSat-2 (RMSD  =  1.5 cm during the two ice growth seasons and for all months of the period of study. The comparison of the altimetric data sets with in situ ice draught measurements during the common flight period shows that the improved Envisat data set (RMSE  =  12–28 cm is as accurate as CryoSat-2 (RMSE  =  15–21 cm and much more accurate than the uncorrected Envisat data set (RMSE  =  178–179 cm. The comparison of the improved Envisat radar freeboard data set is then extended to the rest of the Envisat mission to demonstrate the validity of PP correction from the calibration period. The good agreement between the improved Envisat data set and the in situ ice draught data set (RMSE�

Quantification by aberration corrected (S)TEM of boundaries formed by symmetry breaking phase transformations

Energy Technology Data Exchange (ETDEWEB)

Schryvers, D., E-mail: nick.schryvers@uantwerpen.be [EMAT, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp (Belgium); Salje, E.K.H. [Department of Earth Sciences, University of Cambridge, Cambridge CB2 3EQ (United Kingdom); Nishida, M. [Department of Engineering Sciences for Electronics and Materials, Faculty of Engineering Sciences, Kyushu University, Kasuga, Fukuoka 816-8580 (Japan); De Backer, A. [EMAT, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp (Belgium); Idrissi, H. [EMAT, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp (Belgium); Institute of Mechanics, Materials and Civil Engineering, Université Catholique de Louvain, Place Sainte Barbe, 2, B-1348, Louvain-la-Neuve (Belgium); Van Aert, S. [EMAT, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp (Belgium)

2017-05-15

The present contribution gives a review of recent quantification work of atom displacements, atom site occupations and level of crystallinity in various systems and based on aberration corrected HR(S)TEM images. Depending on the case studied, picometer range precisions for individual distances can be obtained, boundary widths at the unit cell level determined or statistical evolutions of fractions of the ordered areas calculated. In all of these cases, these quantitative measures imply new routes for the applications of the respective materials. - Highlights: • Quantification of picometer displacements at ferroelastic twin boundary in CaTiO{sub 3.} • Quantification of kinks in meandering ferroelectric domain wall in LiNbO{sub 3}. • Quantification of column occupation in anti-phase boundary in Co-Pt. • Quantification of atom displacements at twin boundary in Ni-Ti B19′ martensite.

The New Electron Microscopy: Cells and Molecules in Three Dimensions | Poster

Science.gov (United States)

NCI recently announced the launch of the new National Cryo-Electron Microscopy Facility (NCEF) at the Frederick National Laboratory for Cancer Research (FNLCR). The launch comes while cryo-electron microscopy (cryo-EM) is enjoying the spotlight as a newly emerging, rapidly evolving technology with the potential to revolutionize the field of structural biology. Read more...

Self-correcting electronically scanned pressure sensor

Science.gov (United States)

Gross, C. (Inventor)

1983-01-01

A multiple channel high data rate pressure sensing device is disclosed for use in wind tunnels, spacecraft, airborne, process control, automotive, etc., pressure measurements. Data rates in excess of 100,000 measurements per second are offered with inaccuracies from temperature shifts less than 0.25% (nominal) of full scale over a temperature span of 55 C. The device consists of thirty-two solid state sensors, signal multiplexing electronics to electronically address each sensor, and digital electronic circuitry to automatically correct the inherent thermal shift errors of the pressure sensors and their associated electronics.

Spherical aberrations of human astigmatic corneas.

Science.gov (United States)

Zhao, Huawei; Dai, Guang-Ming; Chen, Li; Weeber, Henk A; Piers, Patricia A

2011-11-01

To evaluate whether the average spherical aberration of human astigmatic corneas is statistically equivalent to human nonastigmatic corneas. Spherical aberrations of 445 astigmatic corneas prior to laser vision correction were retrospectively investigated to determine Zernike coefficients for central corneal areas 6 mm in diameter using CTView (Sarver and Associates). Data were divided into groups according to cylinder power (0.01 to 0.25 diopters [D], 0.26 to 0.75 D, 0.76 to 1.06 D, 1.07 to 1.53 D, 1.54 to 2.00 D, and >2.00 D) and according to age by decade. Spherical aberrations were correlated with age and astigmatic power among groups and the entire population. Statistical analyses were conducted, and P.05 for all tested groups). Mean spherical aberration of astigmatic corneas was not correlated significantly with cylinder power or age (P>.05). Spherical aberrations are similar to those of nonastigmatic corneas, permitting the use of these additional data in the design of aspheric toric intra-ocular lenses. Copyright 2011, SLACK Incorporated.

Cryo-electron microscopy and single molecule fluorescent microscopy detect CD4 receptor induced HIV size expansion prior to cell entry

International Nuclear Information System (INIS)

Pham, Son; Tabarin, Thibault; Garvey, Megan; Pade, Corinna; Rossy, Jérémie; Monaghan, Paul; Hyatt, Alex; Böcking, Till; Leis, Andrew; Gaus, Katharina; Mak, Johnson

2015-01-01

Viruses are often thought to have static structure, and they only remodel after the viruses have entered target cells. Here, we detected a size expansion of virus particles prior to viral entry using cryo-electron microscopy (cryo-EM) and single molecule fluorescence imaging. HIV expanded both under cell-free conditions with soluble receptor CD4 (sCD4) targeting the CD4 binding site on the HIV-1 envelope protein (Env) and when HIV binds to receptor on cellular membrane. We have shown that the HIV Env is needed to facilitate receptor induced virus size expansions, showing that the ‘lynchpin’ for size expansion is highly specific. We demonstrate that the size expansion required maturation of HIV and an internal capsid core with wild type stability, suggesting that different HIV compartments are linked and are involved in remodelling. Our work reveals a previously unknown event in HIV entry, and we propose that this pre-entry priming process enables HIV particles to facilitate the subsequent steps in infection. - Highlights: • Cell free viruses are able to receive external trigger that leads to apparent size expansion. • Virus envelope and CD4 receptor engagement is the lynchpin of virus size expansion. • Internal capsid organisation can influence receptor mediated virus size expansion. • Pre-existing virus-associated lipid membrane in cell free virus can accommodate the receptor mediated virus size expansion.

Cryo-electron microscopy and single molecule fluorescent microscopy detect CD4 receptor induced HIV size expansion prior to cell entry

Energy Technology Data Exchange (ETDEWEB)

Pham, Son [Deakin University, Victoria 3216 (Australia); CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Tabarin, Thibault [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Garvey, Megan; Pade, Corinna [Deakin University, Victoria 3216 (Australia); CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Rossy, Jérémie [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Monaghan, Paul; Hyatt, Alex [CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Böcking, Till [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Leis, Andrew [CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia); Gaus, Katharina, E-mail: k.gaus@unsw.edu.au [ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, New South Wales 3220 (Australia); Mak, Johnson, E-mail: j.mak@deakin.edu.au [Deakin University, Victoria 3216 (Australia); CSIRO Australian Animal Health Laboratory, Victoria 3220 (Australia)

2015-12-15

Viruses are often thought to have static structure, and they only remodel after the viruses have entered target cells. Here, we detected a size expansion of virus particles prior to viral entry using cryo-electron microscopy (cryo-EM) and single molecule fluorescence imaging. HIV expanded both under cell-free conditions with soluble receptor CD4 (sCD4) targeting the CD4 binding site on the HIV-1 envelope protein (Env) and when HIV binds to receptor on cellular membrane. We have shown that the HIV Env is needed to facilitate receptor induced virus size expansions, showing that the ‘lynchpin’ for size expansion is highly specific. We demonstrate that the size expansion required maturation of HIV and an internal capsid core with wild type stability, suggesting that different HIV compartments are linked and are involved in remodelling. Our work reveals a previously unknown event in HIV entry, and we propose that this pre-entry priming process enables HIV particles to facilitate the subsequent steps in infection. - Highlights: • Cell free viruses are able to receive external trigger that leads to apparent size expansion. • Virus envelope and CD4 receptor engagement is the lynchpin of virus size expansion. • Internal capsid organisation can influence receptor mediated virus size expansion. • Pre-existing virus-associated lipid membrane in cell free virus can accommodate the receptor mediated virus size expansion.

Best practices for managing large CryoEM facilities.

Science.gov (United States)

Alewijnse, Bart; Ashton, Alun W; Chambers, Melissa G; Chen, Songye; Cheng, Anchi; Ebrahim, Mark; Eng, Edward T; Hagen, Wim J H; Koster, Abraham J; López, Claudia S; Lukoyanova, Natalya; Ortega, Joaquin; Renault, Ludovic; Reyntjens, Steve; Rice, William J; Scapin, Giovanna; Schrijver, Raymond; Siebert, Alistair; Stagg, Scott M; Grum-Tokars, Valerie; Wright, Elizabeth R; Wu, Shenping; Yu, Zhiheng; Zhou, Z Hong; Carragher, Bridget; Potter, Clinton S

2017-09-01

This paper provides an overview of the discussion and presentations from the Workshop on the Management of Large CryoEM Facilities held at the New York Structural Biology Center, New York, NY on February 6-7, 2017. A major objective of the workshop was to discuss best practices for managing cryoEM facilities. The discussions were largely focused on supporting single-particle methods for cryoEM and topics included: user access, assessing projects, workflow, sample handling, microscopy, data management and processing, and user training. Crown Copyright © 2017. Published by Elsevier Inc. All rights reserved.

On- and off-eye spherical aberration of soft contact lenses and consequent changes of effective lens power.

Science.gov (United States)

Dietze, Holger H; Cox, Michael J

2003-02-01

Soft contact lenses produce a significant level of spherical aberration affecting their power on-eye. A simple model assuming that a thin soft contact lens aligns to the cornea predicts that these effects are similar on-eye and off-eye. The wavefront aberration for 17 eyes and 33 soft contact lenses on-eye was measured with a Shack-Hartmann wavefront sensor. The Zernike coefficients describing the on-eye spherical aberration of the soft contact lens were compared with off-eye ray-tracing results. Paraxial and effective lens power changes were determined. The model predicts the on-eye spherical aberration of soft contact lenses closely. The resulting power change for a +/- 7.00 D spherical soft contact lens is +/- 0.5 D for a 6-mm pupil diameter and +/- 0.1 D for a 3-mm pupil diameter. Power change is negligible for soft contact lenses corrected for off-eye spherical aberration. For thin soft contact lenses, the level of spherical aberration and the consequent power change is similar on-eye and off-eye. Soft contact lenses corrected for spherical aberration in air will be expected to be aberration-free on-eye and produce only negligibly small power changes. For soft contact lenses without aberration correction, for higher levels of ametropia and large pupils, the soft contact lens power should be determined with trial lenses with their power and p value similar to the prescribed lens. The benefit of soft contact lenses corrected for spherical aberration depends on the level of ocular spherical aberration.

Electron fluence correction factors for various materials in clinical electron beams

International Nuclear Information System (INIS)

Olivares, M.; Blois, F. de; Podgorsak, E.B.; Seuntjens, J.P.

2001-01-01

Relative to solid water, electron fluence correction factors at the depth of dose maximum in bone, lung, aluminum, and copper for nominal electron beam energies of 9 MeV and 15 MeV of the Clinac 18 accelerator have been determined experimentally and by Monte Carlo calculation. Thermoluminescent dosimeters were used to measure depth doses in these materials. The measured relative dose at d max in the various materials versus that of solid water, when irradiated with the same number of monitor units, has been used to calculate the ratio of electron fluence for the various materials to that of solid water. The beams of the Clinac 18 were fully characterized using the EGS4/BEAM system. EGSnrc with the relativistic spin option turned on was used to optimize the primary electron energy at the exit window, and to calculate depth doses in the five phantom materials using the optimized phase-space data. Normalizing all depth doses to the dose maximum in solid water stopping power ratio corrected, measured depth doses and calculated depth doses differ by less than ±1% at the depth of dose maximum and by less than 4% elsewhere. Monte Carlo calculated ratios of doses in each material to dose in LiF were used to convert the TLD measurements at the dose maximum into dose at the center of the TLD in the phantom material. Fluence perturbation correction factors for a LiF TLD at the depth of dose maximum deduced from these calculations amount to less than 1% for 0.15 mm thick TLDs in low Z materials and are between 1% and 3% for TLDs in Al and Cu phantoms. Electron fluence ratios of the studied materials relative to solid water vary between 0.83±0.01 and 1.55±0.02 for materials varying in density from 0.27 g/cm3 (lung) to 8.96 g/cm3 (Cu). The difference in electron fluence ratios derived from measurements and calculations ranges from -1.6% to +0.2% at 9 MeV and from -1.9% to +0.2% at 15 MeV and is not significant at the 1σ level. Excluding the data for Cu, electron fluence

Analysis of the 'dilemma effect' in fifth-order deflection aberration

International Nuclear Information System (INIS)

Zhang Xiaobing; Yin Hanchun; Lei Wei; Xue Kunxing; Tong Linsu

1999-01-01

In this paper, the coma of the fifth-order aberration at a large deflection angle has been analyzed by using multipole field theory. The dilemma effect exists in the comas of fifth-order aberration. The dilemma effect, whose value D r is constant and independent of the 10-pole field, is the linear combination of coma aberrations. The coma of the fifth-order aberration is corrected by adjusting the 10-pole field distribution when D r is zero or small. The factors that influence the dilemma effect have been calculated and analyzed

A comparative study of dose distribution of a high-energy electron beam and chromosome aberration frequencies

International Nuclear Information System (INIS)

Matsubara, Sho; Kuwabara, Yuji; Horiuch, Junichi; Suzuki, Soji; Hoshina, Masao; Kato, Tsuguhisa

1986-01-01

Peripheral blood was exposed to a 14 MeV electron beam in a plastic tube set in a test-tube stand immersed in a water tank. The chromosome aberration frequencies induced by irradiation of about 95% of peak dose at a depth of 31 mm were found to be higher in value than those induced at a depth of 17 mm where the peak dose had been determined physically. Three gray of irradiation given to whole blood in the presence of contrast medium gave rise to a slight enhancement of radiation-induced chromosome aberration frequencies in the lymphocytes exposed at a depth of 17 mm, but a slight decrease at 31 mm. (author)

Exploring the atomic structure of 1.8 nm monolayer-protected gold clusters with aberration-corrected STEM

Energy Technology Data Exchange (ETDEWEB)

Liu, Jian; Jian, Nan; Ornelas, Isabel; Pattison, Alexander J. [Nanoscale Physics Research Laboratory, School of Physics and Astronomy, University of Birmingham, Birmingham B15 2TT (United Kingdom); Lahtinen, Tanja; Salorinne, Kirsi [Department of Chemistry, Nanoscience Center, University of Jyväskylä, FI-40014 Jyväskylä (Finland); Häkkinen, Hannu [Department of Chemistry, Nanoscience Center, University of Jyväskylä, FI-40014 Jyväskylä (Finland); Department of Physics, Nanoscience Center, University of Jyväskylä, FI-40014 Jyväskylä (Finland); Palmer, Richard E., E-mail: richardepalmerwork@yahoo.com [Nanoscale Physics Research Laboratory, School of Physics and Astronomy, University of Birmingham, Birmingham B15 2TT (United Kingdom)

2017-05-15

Monolayer-protected (MP) Au clusters present attractive quantum systems with a range of potential applications e.g. in catalysis. Knowledge of the atomic structure is needed to obtain a full understanding of their intriguing physical and chemical properties. Here we employed aberration-corrected scanning transmission electron microscopy (ac-STEM), combined with multislice simulations, to make a round-robin investigation of the atomic structure of chemically synthesised clusters with nominal composition Au{sub 144}(SCH{sub 2}CH{sub 2}Ph){sub 60} provided by two different research groups. The MP Au clusters were “weighed” by the atom counting method, based on their integrated intensities in the high angle annular dark field (HAADF) regime and calibrated exponent of the Z dependence. For atomic structure analysis, we compared experimental images of hundreds of clusters, with atomic resolution, against a variety of structural models. Across the size range 123–151 atoms, only 3% of clusters matched the theoretically predicted Au{sub 144}(SR){sub 60} structure, while a large proportion of the clusters were amorphous (i.e. did not match any model structure). However, a distinct ring-dot feature, characteristic of local icosahedral symmetry, was observed in about 20% of the clusters. - Highlights: • Chemically synthesised gold clusters were “weighed” by atom counting to get true size. • Image simulations show a few percent of clusters have the predicted atomic structure. • But a specific ring-dot feature indicates local icosahedral order in many clusters.

Nonlinear adaptive optics: aberration correction in three photon fluorescence microscopy for mouse brain imaging

Science.gov (United States)

Sinefeld, David; Paudel, Hari P.; Wang, Tianyu; Wang, Mengran; Ouzounov, Dimitre G.; Bifano, Thomas G.; Xu, Chris

2017-02-01

Multiphoton fluorescence microscopy is a well-established technique for deep-tissue imaging with subcellular resolution. Three-photon microscopy (3PM) when combined with long wavelength excitation was shown to allow deeper imaging than two-photon microscopy (2PM) in biological tissues, such as mouse brain, because out-of-focus background light can be further reduced due to the higher order nonlinear excitation. As was demonstrated in 2PM systems, imaging depth and resolution can be improved by aberration correction using adaptive optics (AO) techniques which are based on shaping the scanning beam using a spatial light modulator (SLM). In this way, it is possible to compensate for tissue low order aberration and to some extent, to compensate for tissue scattering. Here, we present a 3PM AO microscopy system for brain imaging. Soliton self-frequency shift is used to create a femtosecond source at 1675 nm and a microelectromechanical (MEMS) SLM serves as the wavefront shaping device. We perturb the 1020 segment SLM using a modified nonlinear version of three-point phase shifting interferometry. The nonlinearity of the fluorescence signal used for feedback ensures that the signal is increasing when the spot size decreases, allowing compensation of phase errors in an iterative optimization process without direct phase measurement. We compare the performance for different orders of nonlinear feedback, showing an exponential growth in signal improvement as the nonlinear order increases. We demonstrate the impact of the method by applying the 3PM AO system for in-vivo mouse brain imaging, showing improvement in signal at 1-mm depth inside the brain.

Three-Dimensional scanning transmission electron microscopy of biological specimens

KAUST Repository

De Jonge, Niels; Sougrat, Rachid; Northan, Brian M.; Pennycook, Stephen J.

2010-01-01

A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM

The effect of pipecol angles for the magnetic electron lens on the aberration coefficients

International Nuclear Information System (INIS)

Al-Khshab, A. M.; Al-Khshab, A. H.

1997-01-01

The symmetric mag etic objective lens of great importance for the electronic microscopes intended for hi g resolution. Such lens is determined, not only by its geometries structure and shape parameters, but also by the influence of the variation of the pole piece angles. the results show that the Objective lens having the pole piece angle of 55 a has a considerable effect on the electron optical Properties. When this pole piece is appropriately and highly saturated, the lens possesses low spherical and chromatic aberration coefficients. This hind of pole piece lens leads to more favourable design than other lenses. (authors). 14 refs., 7 figs.1 table

Cryo-EM Structure Determination Using Segmented Helical Image Reconstruction.

Science.gov (United States)

Fromm, S A; Sachse, C

2016-01-01

Treating helices as single-particle-like segments followed by helical image reconstruction has become the method of choice for high-resolution structure determination of well-ordered helical viruses as well as flexible filaments. In this review, we will illustrate how the combination of latest hardware developments with optimized image processing routines have led to a series of near-atomic resolution structures of helical assemblies. Originally, the treatment of helices as a sequence of segments followed by Fourier-Bessel reconstruction revealed the potential to determine near-atomic resolution structures from helical specimens. In the meantime, real-space image processing of helices in a stack of single particles was developed and enabled the structure determination of specimens that resisted classical Fourier helical reconstruction and also facilitated high-resolution structure determination. Despite the progress in real-space analysis, the combination of Fourier and real-space processing is still commonly used to better estimate the symmetry parameters as the imposition of the correct helical symmetry is essential for high-resolution structure determination. Recent hardware advancement by the introduction of direct electron detectors has significantly enhanced the image quality and together with improved image processing procedures has made segmented helical reconstruction a very productive cryo-EM structure determination method. © 2016 Elsevier Inc. All rights reserved.

Technical and Scientific Progress of the CryoSat-2 Mission

Science.gov (United States)

Shepherd, A.; Wingham, D.; Cullen, R.; Mcmillan, M.; Pagli, C.

2012-12-01

Following the launch of CryoSat-2 in April 2010, we have examined the performance of the CryoSat-2 SAR Interferometer over the continental ice sheets of Antarctica and Greenland, the Artic Ocean, and, for the purposes of calibration, over the oceans. The aim of this exercise has been to provide confirmation of the engineering performance of the radar interferometer, and to provide an initial geophysical validation of the resulting elevation measurements. We have confirmed the engineering performance at system level of the interferometer through performing a sequence of satellite rolls over the oceans, which provide a surface of known behavior and surface gradient. The activity has identified some errors in the SARIN L1b data products presently issued by ESA. Once corrected, the ocean calibration has demonstrated that the interferometer measures across-track surface slopes with a precision of 25 micro-radians and an accuracy of 10 micro-radians, which may be compared with a pre-launch estimation of 100 micro-radians; in short, the engineering performance greatly exceeds its' specification. The elevation measurement over the ice sheets combines the interferometer measurement of across track slope with the range measurement deduced from the SAR echoes. We have examined the performance of the range estimation, and determined the range precision to be 19 cm RMS at 20 Hz. We have examined the retrieval of the phase information over the ice sheets, and found the phase estimates to be robust and little affected by the uncertain ice sheet topography. Based on the calibration of the interferometer, the contribution of the across track slope error is, at 0.4 mm, negligible. While the quantity of data available to us that contains the corrections identified by the interferometer is limited, we have been able to confirm the range precision values from a limited cross-over analysis. Over marine sea ice, we have verified the discrimination of sea ice and ocean lead returns using

CryoSat Level1b SAR/SARin BaselineC: Product Format and Algorithm Improvements

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; Di Giacinto, Andrea; Bouffard, Jerome; Féménias, Pierre; Parrinello, Tommaso

2015-04-01

CryoSat was launched on the 8th April 2010 and is the first European ice mission dedicated to the monitoring of precise changes in the thickness of polar ice sheets and floating sea ice. Cryosat carries an innovative radar altimeter called the Synthetic Aperture Interferometric Altimeter (SIRAL), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach a significantly improved along track resolution with respect to traditional pulse-width limited altimeters. CryoSat is the first altimetry mission operating in SAR mode and continuous improvements in the Level1 Instrument Processing Facility (IPF1) are being identified, tested and validated in order to improve the quality of the Level1b products. The current IPF, Baseline B, was released in operation in February 2012. A reprocessing campaign followed, in order to reprocess the data since July 2010. After more than 2 years of development, the release in operations of Baseline C is expected in the first half of 2015. BaselineC Level1b products will be distributed in an updated format, including for example the attitude information (roll, pitch and yaw) and, for SAR/SARIN, the waveform length doubled with respect to Baseline B. Moreveor, various algorithm improvements have been identified: • a datation bias of about -0.5195 ms will be corrected (SAR/SARIn) • a range bias of about 0.6730 m will be corrected (SAR/SARIn) • a roll bias of 0.1062 deg and a pitch bias of 0.0520 deg • Surface sample stack weighting to filter out the single look echoes acquired at highest look angle, that results in a sharpening of the 20Hz waveforms With the operational release of BaselineC, the second CryoSat reprocessing campaign will be initiated, taking benefit of the upgrade implemented in the IPF1 processing chain but also at IPF2 level. The reprocessing campaign will cover the full Cryosat mission starting on 16th July 2010

Classical Electron Model with QED Corrections

OpenAIRE

Lenk, Ron

2010-01-01

In this article we build a metric for a classical general relativistic electron model with QED corrections. We calculate the stress-energy tensor for the radiative corrections to the Coulomb potential in both the near-field and far-field approximations. We solve the three field equations in both cases by using a perturbative expansion to first order in alpha (the fine-structure constant) while insisting that the usual (+, +, -, -) structure of the stress-energy tensor is maintained. The resul...

Effect of aberration of light in X-ray free electron lasers

International Nuclear Information System (INIS)

Geloni, Gianluca; Kocharyan, Vitali; Saldin, Evgeni

2015-11-01

We discuss the physics of a microbunched electron beam kicked by the dipole field of a corrector magnet by describing the kinematics of coherent undulator radiation after the kick. Particle tracking shows that the electron beam direction changes after the kick, while the orientation of the microbunching wavefront stays unvaried. Therefore, electrons motion and wavefront normal have different directions. Coherent radiation emission in a downstream undulator is expected to be dramatically suppressed as soon as the kick angle becomes larger than the divergence of the output radiation. In fact, according to conventional treatments, coherent radiation is emitted along the normal to the microbunching wavefront. Here we show that kinematics predicts a surprising effect. Namely, a description of coherent undulator radiation in the laboratory frame yields the radical notion that, due light aberration, strong coherent radiation is produced along the direction of the kick. We hold a recent FEL study made at the LCLS as a direct experimental evidence that coherent undulator radiation can be kicked by an angle of about five times the rms radiation divergence without suppression. We put forward our kinematical description of this experiment.

Differential algebraic method for arbitrary order curvilinear-axis combined geometric-chromatic aberration analysis

International Nuclear Information System (INIS)

Cheng Min; Tang Tiantong; Lu Yilong; Yao Zhenhua

2003-01-01

The principle of differential algebra is applied to analyse and calculate arbitrary order curvilinear-axis combined geometric-chromatic aberrations of electron optical systems. Expressions of differential algebraic form of high order combined aberrations are obtained and arbitrary order combined aberrations can be calculated numerically. As an example, a typical wide electron beam focusing system with curved optical axes named magnetic immersion lens has been studied. All the second-order and third-order combined geometric-chromatic aberrations of the lens have been calculated, and the patterns of the corresponding geometric aberrations and combined aberrations have been given as well

Aberration characteristics of immersion lenses for LVSEM

International Nuclear Information System (INIS)

Khursheed, Anjam

2002-01-01

This paper investigates the on-axis aberration characteristics of various immersion objective lenses for low voltage scanning electron microscopy (LVSEM). A simple aperture lens model is used to generate smooth axial field distributions. The simulation results show that mixed field electric-magnetic immersion lenses are predicted to have between 1.5 and 2 times smaller aberration limited probe diameters than their pure-field counterparts. At a landing energy of 1 keV, mixed field immersion lenses operating at the vacuum electrical field breakdown limit are predicted to have on-axis aberration coefficients between 50 and 60 μm, yielding an ultimate image resolution of below 1 nm. These aberrations lie in the same range as those for LVSEM systems that employ aberration correctors

Fifth-order canonical geometric aberration analysis of electrostatic round lenses

CERN Document Server

Liu Zhi Xiong

2002-01-01

In this paper the fifth-order canonical geometric aberration patterns are analyzed and a numerical example is given on the basis of the analytical expressions of fifth-order aberration coefficients derived in the present work. The fifth-order spherical aberration, astigmatism and field curvature, and distortion are similar to the third-order ones and the fifth-order coma is slightly different. Besides, there are two more aberrations which do not exist in the third-order aberration: they are peanut aberration and elliptical coma in accordance with their shapes. In the numerical example, by using a cross-check of the calculated coefficients with those computed through the differential algebraic method, it has been verified that all the expressions are correct and the computational results are reliable with high precision.

Transmission electron microscopy of amyloid fibrils.

Science.gov (United States)

Gras, Sally L; Waddington, Lynne J; Goldie, Kenneth N

2011-01-01

Transmission Electron Microscopy of negatively stained and cryo-prepared specimens allows amyloid fibrils to be visualised at high resolution in a dried or a hydrated state, and is an essential method for characterising the morphology of fibrils and pre-fibrillar species. We outline the key steps involved in the preparation and observation of samples using negative staining and cryo-electron preservation. We also discuss methods to measure fibril characteristics, such as fibril width, from electron micrographs.

Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry.

Science.gov (United States)

Schwertner, M; Booth, M J; Neil, M A A; Wilson, T

2004-01-01

Confocal or multiphoton microscopes, which deliver optical sections and three-dimensional (3D) images of thick specimens, are widely used in biology. These techniques, however, are sensitive to aberrations that may originate from the refractive index structure of the specimen itself. The aberrations cause reduced signal intensity and the 3D resolution of the instrument is compromised. It has been suggested to correct for aberrations in confocal microscopes using adaptive optics. In order to define the design specifications for such adaptive optics systems, one has to know the amount of aberrations present for typical applications such as with biological samples. We have built a phase stepping interferometer microscope that directly measures the aberration of the wavefront. The modal content of the wavefront is extracted by employing Zernike mode decomposition. Results for typical biological specimens are presented. It was found for all samples investigated that higher order Zernike modes give only a small contribution to the overall aberration. Therefore, these higher order modes can be neglected in future adaptive optics sensing and correction schemes implemented into confocal or multiphoton microscopes, leading to more efficient designs.

CryoSat-2 SIRAL Calibration: Strategy, Application and Results

Science.gov (United States)

Parrinello, T.; Fornari, M.; Bouzinac, C.; Scagliola, M.; Tagliani, N.

2012-04-01

The main payload of CryoSat-2 is a Ku band pulsewidth limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. This allows to reach an along track resolution of about 250 meters which is an important improvement over traditional pulse-width limited altimeters. Due to the fact that SIRAL is a phase coherent pulse-width limited radar altimeter, a proper calibration approach has been developed. In fact, not only the corrections for transfer function amplitude with respect to frequency, gain and instrument path delay have to be computed but it is also needed to provide corrections for transfer function phase with respect to frequency and AGC setting as well as the phase variation across bursts of pulses. As a consequence, SIRAL performs regularly four types of calibrations: (1) CAL1 in order to calibrate the internal path delay and peak power variation, (2) CAL2 in order to compensate the instrument transfer function, (3) CAL4 to calibrate the interferometer and (4) AutoCal, a specific sequence in order to calibrate the gain and phase difference for each AGC setting. Commissioning phase results (April-December 2010) revealed high stability of the instrument, which made possible to reduce the calibration frequency during Operations. Internal calibration data are processed on ground by the CryoSat-2 Instrument Processing Facility (IPF1) and then applied to the science data. In this poster we will describe as first the calibration strategy and then how the four different types of calibration are applied to science data. Moreover the calibration results over almost 2 years of mission will be presented, analyzing their temporal evolution in order to highlight the stability of the instrument over its life.

Understanding the structure of nanocatalysts with high resolution scanning/transmission electron microscopy

International Nuclear Information System (INIS)

Francis, L D; Rivas, J; José-Yacamán, M

2014-01-01

Nanomaterials including nanoparticles, nanowires and nanotubes play an important role in heterogeneous catalysis. Thanks to the rapid improvement of the electron microscopic techniques and with the advent of aberration corrected electron microscopy as well as theoretical methodologies, the potential effects induced by nanocatalysts are better understood than before by unravelling their atomic structure. A brief introduction to advanced electron microscopic techniques namely aberration corrected scanning transmission electron microscopy (Cs-STEM) is presented and subsequently two examples of nanocatalysts are considered in the present review. The first example will focus on the study of bimetallic/core-shell nanoalloys. In heterogeneous catalysis, catalysts containing two or more metals might show significantly different catalytic properties compared to the parent metals and thus are widely utilized in several catalytic reactions. Atom-by-atom insights of the nanoalloy based catalysts ex: Au-Pd will be described in the present review using a combination of advanced electron microscopic and spectroscopic techniques. A related example on the understanding of bimetallic clusters by HAADF-STEM will also be presented in addition to nanoparticles. In the second case understanding the structure of transition metal chalcogenide based nanocatalysts by HRTEM and aberration corrected STEM, for the case of MoS 2 will be discussed. MoS 2 -based catalysts serve as model catalysts and are employed in the hydrodesulphurisations (HDS) reactions in the removal of sulphur from gasoline and related petrochemical products. They have been studied in various forms including nanowires, nanotubes and nanoplates. Their structure, atomic insights and as a consequence elucidation of their corresponding catalytic activity are thus important

Structure of a headful DNA-packaging bacterial virus at 2.9 Å resolution by electron cryo-microscopy.

Science.gov (United States)

Zhao, Haiyan; Li, Kunpeng; Lynn, Anna Y; Aron, Keith E; Yu, Guimei; Jiang, Wen; Tang, Liang

2017-04-04

The enormous prevalence of tailed DNA bacteriophages on this planet is enabled by highly efficient self-assembly of hundreds of protein subunits into highly stable capsids. These capsids can stand with an internal pressure as high as ∼50 atmospheres as a result of the phage DNA-packaging process. Here we report the complete atomic model of the headful DNA-packaging bacteriophage Sf6 at 2.9 Å resolution determined by electron cryo-microscopy. The structure reveals the DNA-inflated, tensed state of a robust protein shell assembled via noncovalent interactions. Remarkable global conformational polymorphism of capsid proteins, a network formed by extended N arms, mortise-and-tenon-like intercapsomer joints, and abundant β-sheet-like mainchain:mainchain intermolecular interactions, confers significant strength yet also flexibility required for capsid assembly and DNA packaging. Differential formations of the hexon and penton are mediated by a drastic α-helix-to-β-strand structural transition. The assembly scheme revealed here may be common among tailed DNA phages and herpesviruses.

Electron tomography and cryo-SEM characterization reveals novel ultrastructural features of host-parasite interaction during Chlamydia abortus infection.

Science.gov (United States)

Wilkat, M; Herdoiza, E; Forsbach-Birk, V; Walther, P; Essig, A

2014-08-01

Chlamydia (C.) abortus is a widely spread pathogen among ruminants that can be transmitted to women during pregnancy leading to severe systemic infection with consecutive abortion. As a member of the Chlamydiaceae, C. abortus shares the characteristic feature of an obligate intracellular biphasic developmental cycle with two morphological forms including elementary bodies (EBs) and reticulate bodies (RBs). In contrast to other chlamydial species, C. abortus ultrastructure has not been investigated yet. To do so, samples were fixed by high-pressure freezing and processed by different electron microscopic methods. Freeze-substituted samples were analysed by transmission electron microscopy, scanning transmission electron microscopical tomography and immuno-electron microscopy, and freeze-fractured samples were analysed by cryo-scanning electron microscopy. Here, we present three ultrastructural features of C. abortus that have not been reported up to now. Firstly, the morphological evidence that C. abortus is equipped with the type three secretion system. Secondly, the accumulation and even coating of whole inclusion bodies by membrane complexes consisting of multiple closely adjacent membranes which seems to be a C. abortus specific feature. Thirdly, the formation of small vesicles in the periplasmic space of RBs in the second half of the developmental cycle. Concerning the time point of their formation and the fact that they harbour chlamydial components, these vesicles might be morphological correlates of an intermediate step during the process of redifferentiation of RBs into EBs. As this feature has also been shown for C. trachomatis and C. pneumoniae, it might be a common characteristic of the family of Chlamydiaceae.

A novel low energy electron microscope for DNA sequencing and surface analysis

Energy Technology Data Exchange (ETDEWEB)

Mankos, M., E-mail: marian@electronoptica.com [Electron Optica Inc., 1000 Elwell Court #110, Palo Alto, CA 94303 (United States); Shadman, K. [Electron Optica Inc., 1000 Elwell Court #110, Palo Alto, CA 94303 (United States); Persson, H.H.J. [Stanford Genome Technology Center, Stanford University School of Medicine, 855 California Avenue, Palo Alto, CA 94304 (United States); N’Diaye, A.T. [Electron Optica Inc., 1000 Elwell Court #110, Palo Alto, CA 94303 (United States); NCEM, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States); Schmid, A.K. [NCEM, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States); Davis, R.W. [Stanford Genome Technology Center, Stanford University School of Medicine, 855 California Avenue, Palo Alto, CA 94304 (United States)

2014-10-15

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel technique that is directed towards imaging nanostructures and surfaces with sub-nanometer resolution. The technique combines a monochromator, a mirror aberration corrector, an energy filter, and dual beam illumination in a single instrument. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. Simulation results predict that the novel aberration corrector design will eliminate the second rank chromatic and third and fifth order spherical aberrations, thereby improving the resolution into the sub-nanometer regime at landing energies as low as one hundred electron-Volts. The energy filter produces a beam that can extract detailed information about the chemical composition and local electronic states of non-periodic objects such as nanoparticles, interfaces, defects, and macromolecules. The dual flood illumination eliminates charging effects that are generated when a conventional LEEM is used to image insulating specimens. A potential application for MAD-LEEM is in DNA sequencing, which requires high resolution to distinguish the individual bases and high speed to reduce the cost. The MAD-LEEM approach images the DNA with low electron impact energies, which provides nucleobase contrast mechanisms without organometallic labels. Furthermore, the micron-size field of view when combined with imaging on the fly provides long read lengths, thereby reducing the demand on assembling the sequence. Experimental results from bulk specimens with immobilized single-base oligonucleotides demonstrate that base specific contrast is available with reflected, photo-emitted, and Auger electrons. Image contrast simulations of model rectangular features mimicking the individual nucleotides in a DNA strand have been developed to translate measurements of contrast on bulk DNA to the detectability of

Cryo-Scanning Electron Microscopy (SEM) and Scanning Transmission Electron Microscopy (STEM)-in-SEM for Bio- and Organo-Mineral Interface Characterization in the Environment.

Science.gov (United States)

Wille, Guillaume; Hellal, Jennifer; Ollivier, Patrick; Richard, Annie; Burel, Agnes; Jolly, Louis; Crampon, Marc; Michel, Caroline

2017-12-01

Understanding biofilm interactions with surrounding substratum and pollutants/particles can benefit from the application of existing microscopy tools. Using the example of biofilm interactions with zero-valent iron nanoparticles (nZVI), this study aims to apply various approaches in biofilm preparation and labeling for fluorescent or electron microscopy and energy dispersive X-ray spectrometry (EDS) microanalysis for accurate observations. According to the targeted microscopy method, biofilms were sampled as flocs or attached biofilm, submitted to labeling using 4',6-diamidino-2-phenylindol, lectins PNA and ConA coupled to fluorescent dye or gold nanoparticles, and prepared for observation (fixation, cross-section, freezing, ultramicrotomy). Fluorescent microscopy revealed that nZVI were embedded in the biofilm structure as aggregates but the resolution was insufficient to observe individual nZVI. Cryo-scanning electron microscopy (SEM) observations showed nZVI aggregates close to bacteria, but it was not possible to confirm direct interactions between nZVI and cell membranes. Scanning transmission electron microscopy in the SEM (STEM-in-SEM) showed that nZVI aggregates could enter the biofilm to a depth of 7-11 µm. Bacteria were surrounded by a ring of extracellular polymeric substances (EPS) preventing direct nZVI/membrane interactions. STEM/EDS mapping revealed a co-localization of nZVI aggregates with lectins suggesting a potential role of EPS in nZVI embedding. Thus, the combination of divergent microscopy approaches is a good approach to better understand and characterize biofilm/metal interactions.

Zernike phase contrast cryo-electron tomography of sodium-driven flagellar hook-basal bodies from Vibrio alginolyticus.

Science.gov (United States)

Hosogi, Naoki; Shigematsu, Hideki; Terashima, Hiroyuki; Homma, Michio; Nagayama, Kuniaki

2011-01-01

Vibrio alginolyticus use flagella to swim. A flagellum consists of a filament, hook and basal body. The basal body is made up of a rod and several ring structures. This study investigates the structure of the T ring which is a unique component of the V. alginolyticus sodium ion-driven flagellar basal body. Using Zernike phase contrast (ZPC) cryo-electron tomography, we compared the 3D structures of purified hook-basal bodies (HBB) from a wild-type strain (KK148) and a deletion mutant lacking MotX and MotY (TH3), which are thought to form the T ring. ZPC images of HBBs had highly improved signal-to-noise ratio compared to conventional phase contrast images. We observed the outline of the HBBs from strains KK148 and TH3, and the TH3 mutant was missing its T ring. In the wild-type strain, the T ring was beneath the LP ring and seemed to form a ring shape with diameter of 32 nm. Copyright © 2010 Elsevier Inc. All rights reserved.

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

Science.gov (United States)

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

2015-12-01

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

Fingerprint-based structure retrieval using electron density.

Science.gov (United States)

Yin, Shuangye; Dokholyan, Nikolay V

2011-03-01

We present a computational approach that can quickly search a large protein structural database to identify structures that fit a given electron density, such as determined by cryo-electron microscopy. We use geometric invariants (fingerprints) constructed using 3D Zernike moments to describe the electron density, and reduce the problem of fitting of the structure to the electron density to simple fingerprint comparison. Using this approach, we are able to screen the entire Protein Data Bank and identify structures that fit two experimental electron densities determined by cryo-electron microscopy. Copyright © 2010 Wiley-Liss, Inc.

eV-TEM: Transmission electron microscopy in a low energy cathode lens instrument

Energy Technology Data Exchange (ETDEWEB)

Geelen, Daniël, E-mail: geelen@physics.leidenuniv.nl [Huygens-Kamerlingh Onnes Laboratory, Leiden Institute of Physics, Leiden University, P.O. Box 9504, 2300 RA Leiden (Netherlands); Thete, Aniket [Huygens-Kamerlingh Onnes Laboratory, Leiden Institute of Physics, Leiden University, P.O. Box 9504, 2300 RA Leiden (Netherlands); Schaff, Oliver; Kaiser, Alexander [SPECS GmbH, Voltastrasse 5, D-13355 Berlin (Germany); Molen, Sense Jan van der [Huygens-Kamerlingh Onnes Laboratory, Leiden Institute of Physics, Leiden University, P.O. Box 9504, 2300 RA Leiden (Netherlands); Tromp, Rudolf [IBM T.J. Watson Research Center, 1101 Kitchawan Road, P.O. Box 218, Yorktown Heights, NY 10598 (United States)

2015-12-15

We are developing a transmission electron microscope that operates at extremely low electron energies, 0–40 eV. We call this technique eV-TEM. Its feasibility is based on the fact that at very low electron energies the number of energy loss pathways decreases. Hence, the electron inelastic mean free path increases dramatically. eV-TEM will enable us to study elastic and inelastic interactions of electrons with thin samples. With the recent development of aberration correction in cathode lens instruments, a spatial resolution of a few nm appears within range, even for these very low electron energies. Such resolution will be highly relevant to study biological samples such as proteins and cell membranes. The low electron energies minimize adverse effects due to radiation damage. - Highlights: • We present a new way of performing low energy transmission electron microscopy in an aberration corrected LEEM/PEEM instrument. • We show a proof of principle where we measure transmitted electrons through a suspended graphene monolayer with a preliminary setup. • We present an improved setup design that provides better control of the incident electron beam.

Massively parallel unsupervised single-particle cryo-EM data clustering via statistical manifold learning.

Directory of Open Access Journals (Sweden)

Jiayi Wu

Full Text Available Structural heterogeneity in single-particle cryo-electron microscopy (cryo-EM data represents a major challenge for high-resolution structure determination. Unsupervised classification may serve as the first step in the assessment of structural heterogeneity. However, traditional algorithms for unsupervised classification, such as K-means clustering and maximum likelihood optimization, may classify images into wrong classes with decreasing signal-to-noise-ratio (SNR in the image data, yet demand increased computational costs. Overcoming these limitations requires further development of clustering algorithms for high-performance cryo-EM data processing. Here we introduce an unsupervised single-particle clustering algorithm derived from a statistical manifold learning framework called generative topographic mapping (GTM. We show that unsupervised GTM clustering improves classification accuracy by about 40% in the absence of input references for data with lower SNRs. Applications to several experimental datasets suggest that our algorithm can detect subtle structural differences among classes via a hierarchical clustering strategy. After code optimization over a high-performance computing (HPC environment, our software implementation was able to generate thousands of reference-free class averages within hours in a massively parallel fashion, which allows a significant improvement on ab initio 3D reconstruction and assists in the computational purification of homogeneous datasets for high-resolution visualization.

Massively parallel unsupervised single-particle cryo-EM data clustering via statistical manifold learning.

Science.gov (United States)

Wu, Jiayi; Ma, Yong-Bei; Congdon, Charles; Brett, Bevin; Chen, Shuobing; Xu, Yaofang; Ouyang, Qi; Mao, Youdong

2017-01-01

Structural heterogeneity in single-particle cryo-electron microscopy (cryo-EM) data represents a major challenge for high-resolution structure determination. Unsupervised classification may serve as the first step in the assessment of structural heterogeneity. However, traditional algorithms for unsupervised classification, such as K-means clustering and maximum likelihood optimization, may classify images into wrong classes with decreasing signal-to-noise-ratio (SNR) in the image data, yet demand increased computational costs. Overcoming these limitations requires further development of clustering algorithms for high-performance cryo-EM data processing. Here we introduce an unsupervised single-particle clustering algorithm derived from a statistical manifold learning framework called generative topographic mapping (GTM). We show that unsupervised GTM clustering improves classification accuracy by about 40% in the absence of input references for data with lower SNRs. Applications to several experimental datasets suggest that our algorithm can detect subtle structural differences among classes via a hierarchical clustering strategy. After code optimization over a high-performance computing (HPC) environment, our software implementation was able to generate thousands of reference-free class averages within hours in a massively parallel fashion, which allows a significant improvement on ab initio 3D reconstruction and assists in the computational purification of homogeneous datasets for high-resolution visualization.

Changes in higher order aberrations after wavefront-guided PRK for correction of low to moderate myopia and myopic astigmatism: two-year follow-up.

Science.gov (United States)

Wigledowska-Promienska, D; Zawojska, I

2007-01-01

To assess efficacy, safety, and changes in higher order aberrations after wavefront-guided photorefractive keratectomy (PRK) in comparison with conventional PRK for low to moderate myopia with myopic astigmatism using a WASCA Workstation with the MEL 70 G-Scan excimer laser. A total of 126 myopic or myopic-astigmatic eyes of 112 patients were included in this retrospective study. Patients were divided into two groups: Group 1, the study group; and Group 2, the control group. Group 1 consisted of 78 eyes treated with wavefront-guided PRK. Group 2 consisted of 48 eyes treated with spherocylindrical conventional PRK. Two years postoperatively, in Group 1, 5% of eyes achieved an uncorrected visual acuity (UCVA) of 0.05; 69% achieved a UCVA of 0.00; 18% of eyes experienced enhanced visual acuity of -0.18 and 8% of -0.30. In Group 2, 8% of eyes achieved a UCVA of 0.1; 25% achieved a UCVA of 0.05; and 67% achieved a UCVA of 0.00 according to logMAR calculation method. Total higher-order root-mean square increased by a factor 1.18 for Group 1 and 1.6 for Group 2. There was a significant increase of coma by a factor 1.74 in Group 2 and spherical aberration by a factor 2.09 in Group 1 and 3.56 in Group 2. The data support the safety and effectiveness of the wavefront-guided PRK using a WASCA Workstation for correction of low to moderate refractive errors. This method reduced the number of higher order aberrations induced by excimer laser surgery and improved uncorrected and spectacle-corrected visual acuity when compared to conventional PRK.

Experimental demonstration of passive acoustic imaging in the human skull cavity using CT-based aberration corrections.

Science.gov (United States)

Jones, Ryan M; O'Reilly, Meaghan A; Hynynen, Kullervo

2015-07-01

Experimentally verify a previously described technique for performing passive acoustic imaging through an intact human skull using noninvasive, computed tomography (CT)-based aberration corrections Jones et al. [Phys. Med. Biol. 58, 4981-5005 (2013)]. A sparse hemispherical receiver array (30 cm diameter) consisting of 128 piezoceramic discs (2.5 mm diameter, 612 kHz center frequency) was used to passively listen through ex vivo human skullcaps (n = 4) to acoustic emissions from a narrow-band fixed source (1 mm diameter, 516 kHz center frequency) and from ultrasound-stimulated (5 cycle bursts, 1 Hz pulse repetition frequency, estimated in situ peak negative pressure 0.11-0.33 MPa, 306 kHz driving frequency) Definity™ microbubbles flowing through a thin-walled tube phantom. Initial in vivo feasibility testing of the method was performed. The performance of the method was assessed through comparisons to images generated without skull corrections, with invasive source-based corrections, and with water-path control images. For source locations at least 25 mm from the inner skull surface, the modified reconstruction algorithm successfully restored a single focus within the skull cavity at a location within 1.25 mm from the true position of the narrow-band source. The results obtained from imaging single bubbles are in good agreement with numerical simulations of point source emitters and the authors' previous experimental measurements using source-based skull corrections O'Reilly et al. [IEEE Trans. Biomed. Eng. 61, 1285-1294 (2014)]. In a rat model, microbubble activity was mapped through an intact human skull at pressure levels below and above the threshold for focused ultrasound-induced blood-brain barrier opening. During bursts that led to coherent bubble activity, the location of maximum intensity in images generated with CT-based skull corrections was found to deviate by less than 1 mm, on average, from the position obtained using source-based corrections. Taken

Airborne campaigns for CryoSat prelaunch calibration and validation

DEFF Research Database (Denmark)

Skourup, Henriette; Hanson, Susanne; Hvidegaard, Sine Munk

2011-01-01

After the successful launch of CryoSat-2 in April 2010, the first direct validation campaign of the satellite is planned for spring 2011. DTU Space has been involved in ESA’s CryoSat Validation Experiment (CryoVEx) with airborne activities since 2003. To validate the prelaunch performance...... of the CryoSat radar altimeter (SIRAL), an airborne version of the SIRAL altimeter (ASIRAS) has been flown together with a laser scanner in 2006 and 2008. Of particular interest is to study the penetration depth of the radar altimeter over both land- and sea ice. This can be done by comparing the radar...... and laser measurements with in situ observations. Here, an overview of the prelaunch airborne campaigns is given, together with results of the ASIRAS performance over land- and sea ice. The observations, used in this study, are obtained from the Greenland ice sheet and from both multiyear and first year sea...

Monitoring of Multilayered Bacterial Biofilm Morphology by Cryo-SEM for Raman Spectroscopy Measurements

Czech Academy of Sciences Publication Activity Database

Hrubanová, Kamila; Bernatová, Silvie; Samek, Ota; Šerý, Mojmír; Zemánek, Pavel; Nebesářová, Jana; Růžička, F.; Krzyžánek, Vladislav

2015-01-01

Roč. 21, S3 (2015), s. 187-188 ISSN 1431-9276 R&D Projects: GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01; GA ČR(CZ) GA14-20012S Institutional support: RVO:68081731 ; RVO:60077344 Keywords : multilayered bacterial biofilm * morphology by Cryo-SEM * Raman spectroscopy Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.730, year: 2015

Thon rings from amorphous ice and implications of beam-induced Brownian motion in single particle electron cryo-microscopy.

Science.gov (United States)

McMullan, G; Vinothkumar, K R; Henderson, R

2015-11-01

We have recorded dose-fractionated electron cryo-microscope images of thin films of pure flash-frozen amorphous ice and pre-irradiated amorphous carbon on a Falcon II direct electron detector using 300 keV electrons. We observe Thon rings [1] in both the power spectrum of the summed frames and the sum of power spectra from the individual frames. The Thon rings from amorphous carbon images are always more visible in the power spectrum of the summed frames whereas those of amorphous ice are more visible in the sum of power spectra from the individual frames. This difference indicates that while pre-irradiated carbon behaves like a solid during the exposure, amorphous ice behaves like a fluid with the individual water molecules undergoing beam-induced motion. Using the measured variation in the power spectra amplitude with number of electrons per image we deduce that water molecules are randomly displaced by a mean squared distance of ∼1.1 Å(2) for every incident 300 keV e(-)/Å(2). The induced motion leads to an optimal exposure with 300 keV electrons of 4.0 e(-)/Å(2) per image with which to observe Thon rings centred around the strong 3.7 Å scattering peak from amorphous ice. The beam-induced movement of the water molecules generates pseudo-Brownian motion of embedded macromolecules. The resulting blurring of single particle images contributes an additional term, on top of that from radiation damage, to the minimum achievable B-factor for macromolecular structure determination. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Defining the limits and reliability of rigid-body fitting in cryo-EM maps using multi-scale image pyramids

NARCIS (Netherlands)

van Zundert, G. C P; Bonvin, A. M J J

Cryo-electron microscopy provides fascinating structural insight into large macromolecular machines at increasing detail. Despite significant advances in the field, the resolution of the resulting three-dimensional images is still typically insufficient for . de novo model building. To bridge the

Double aberration-corrected TEM/STEM of tungstated zirconia nanocatalysts for the synthesis of paracetamol

International Nuclear Information System (INIS)

Yoshida, K; Boyes, E D; Gai, P L; Shiju, N R; Brown, D R

2010-01-01

We report highly active tungstated zirconia nanocatalysts for the synthesis of paracetamol by Beckmann rearrangement of 4-hydroxyacetophenone oxime. Double aberration-corrected (2AC)-TEM/STEM studies were performed in a JEOL 2200FS FEG TEM/STEM at the 1 Angstrom (1 A = 0.1 nanometer) level. Observations at close to zero defocus were carried out using the AC-TEM as well as AC-STEM including high angle annular dark field (HAADF) imaging, from the same areas of the catalyst crystallites. The studies from the same areas have revealed the location and the nanostructure of the polytungstate species (clusters) and the nanograins of zirconia. The AC (S)TEM was crucial to observe the nanostructure and location of polytungstate clusters on the zirconia grains. Polytungstate clusters as small as 0.5 nm have been identified using the HAADF-STEM. The nanostructures of the catalyst and the W surface density have been correlated with paracetamol reaction studies. The results demonstrate the nature of active sites and high activity of the tungstated zirconia nanocatalyst, which is an environmentally clean alternative to the current homogeneous process.

Double aberration-corrected TEM/STEM of tungstated zirconia nanocatalysts for the synthesis of paracetamol

Energy Technology Data Exchange (ETDEWEB)

Yoshida, K; Boyes, E D; Gai, P L [York JEOL Nanocentre (United Kingdom); Shiju, N R; Brown, D R, E-mail: pgb500@york.ac.u [Department of Chemical and Biological Sciences, University of Huddersfield, Huddersfield, HD1 3DH (United Kingdom)

2010-07-01

We report highly active tungstated zirconia nanocatalysts for the synthesis of paracetamol by Beckmann rearrangement of 4-hydroxyacetophenone oxime. Double aberration-corrected (2AC)-TEM/STEM studies were performed in a JEOL 2200FS FEG TEM/STEM at the 1 Angstrom (1 A = 0.1 nanometer) level. Observations at close to zero defocus were carried out using the AC-TEM as well as AC-STEM including high angle annular dark field (HAADF) imaging, from the same areas of the catalyst crystallites. The studies from the same areas have revealed the location and the nanostructure of the polytungstate species (clusters) and the nanograins of zirconia. The AC (S)TEM was crucial to observe the nanostructure and location of polytungstate clusters on the zirconia grains. Polytungstate clusters as small as 0.5 nm have been identified using the HAADF-STEM. The nanostructures of the catalyst and the W surface density have been correlated with paracetamol reaction studies. The results demonstrate the nature of active sites and high activity of the tungstated zirconia nanocatalyst, which is an environmentally clean alternative to the current homogeneous process.

Chromosome painting analysis of radiation-induced aberrant cell clones in the mouse

International Nuclear Information System (INIS)

Spruill, M.D.; Nath, J.; Tucker, J.D.

1997-01-01

In a study of the persistence of radiation-induced translocations over the life span of the mouse, we observed a number of clonal cells in peripheral blood lymphocytes. The presence of clones caused the mean frequency of aberrations at various time points to be elevated which interfered with biodosimetry. For this reason, we have corrected our data for the presence of clones. Mice were given an acute dose of 0, 1, 2, 3 or 4 Gy 137 Cs at 8 weeks of age. Aberrations were measured by painting chromosomes 2 and 8 and cells were examined for clones at 3 months and every 3 months thereafter until 21 months. Clones were identified by comparing the color photographic slides of all abnormal cells from each animal. Determination of clonality was made on the basis of similar breakpoint locations or the presence of other identifying characteristics such as unusual aberrations. To correct the frequency of translocations for the presence of clones, each clone, regardless of how many cells it contained, was counted only once. This reflects the original aberration frequency since each clone originated as only one cell. Among mice exposed to 4 Gy, the mean frequencies of aberrant cell clones ranged from 3-29% of the total number of metaphase cells scored with the highest frequency being 1 year post exposure. 32-70% of reciprocal and 19-92% of non-reciprocal translocations were clonal. A dose response relationship for clones was evident until 21 months when the unexposed animals exhibited a mean frequency of aberrant cell clones >10% of the total number of cells scored. Almost 75% of reciprocal and 95% of non-reciprocal translocations in these unexposed control animals were of clonal origin. Correction for clonal expansion greatly reduced the means and their standard errors at most time points where clonal expansion was prevalent. The biodosimetry was much improved suggesting that correction is beneficial in long-term studies

Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography.

Science.gov (United States)

Bharat, Tanmay A M; Noda, Takeshi; Riches, James D; Kraehling, Verena; Kolesnikova, Larissa; Becker, Stephan; Kawaoka, Yoshihiro; Briggs, John A G

2012-03-13

Ebola virus is a highly pathogenic filovirus causing severe hemorrhagic fever with high mortality rates. It assembles heterogenous, filamentous, enveloped virus particles containing a negative-sense, single-stranded RNA genome packaged within a helical nucleocapsid (NC). We have used cryo-electron microscopy and tomography to visualize Ebola virus particles, as well as Ebola virus-like particles, in three dimensions in a near-native state. The NC within the virion forms a left-handed helix with an inner nucleoprotein layer decorated with protruding arms composed of VP24 and VP35. A comparison with the closely related Marburg virus shows that the N-terminal region of nucleoprotein defines the inner diameter of the Ebola virus NC, whereas the RNA genome defines its length. Binding of the nucleoprotein to RNA can assemble a loosely coiled NC-like structure; the loose coil can be condensed by binding of the viral matrix protein VP40 to the C terminus of the nucleoprotein, and rigidified by binding of VP24 and VP35 to alternate copies of the nucleoprotein. Four proteins (NP, VP24, VP35, and VP40) are necessary and sufficient to mediate assembly of an NC with structure, symmetry, variability, and flexibility indistinguishable from that in Ebola virus particles released from infected cells. Together these data provide a structural and architectural description of Ebola virus and define the roles of viral proteins in its structure and assembly.

Synthesis and characterization of a cryo gel starting from chitosan and their study as adsorbent of Cu (II) ions in aqueous solution

International Nuclear Information System (INIS)

Arcos A, A. J.

2012-01-01

The present work consisted in the synthesis of cryo gels from chitosan, which were cross-linked with ethylene glycol diglycidyl ether to obtain a stable, porous and resistant material to the interaction with aqueous solutions, in order that can be used in metal sorption processes. The performed synthesis consisted in a aqueous chitosan gel preparation crosslinked with ethylene glycol diglycidyl ether (Edge), evaluating two levels of crosslinking (1 and 2%); later to obtain the cryo gels, the aqueous gels were subjected to an ice segregation induced self-assembly process (ISISA), considering two levels of speed of materials formation. The obtained materials were characterized first determining their solubility in aqueous solutions and to the pair was determined the degree of swelling of the materials. Later, were used techniques like: the sweeping electronic microscopy to be able to observe the morphology of the obtained cryo gels, X-ray diffraction to know the crystallinity of the same, zeta potential with the aim of know the electrical potential in the materials, specific area determination, Fourier transformation infrared spectroscopy (Ftir) and X-ray photoelectrons spectroscopy (XP S), to know the functional groups and chemical interactions of the present elements in the materials before and after the copper ions sorption. The evaluation of the cryo gels as sorbents was performed obtaining the sorption kinetics for the Cu (II) ion at 25 and 30 grades C, and adjusting the experimental data obtained to the Lagergren and Ho kinetic models, as well as was performed adsorption isotherms at 25 and 35 grades C for the copper ion, adjusting the obtained data to the Langmuir, Freundlich and Langmuir-Freundlich models. In general, it was observed that the obtained materials show a macroporous structure, which presents and uniform swelling without significant deformations. Furthermore, was observed that the speed of formation of the materials affects the pores size and the

Robust image alignment for cryogenic transmission electron microscopy.

Science.gov (United States)

McLeod, Robert A; Kowal, Julia; Ringler, Philippe; Stahlberg, Henning

2017-03-01

Cryo-electron microscopy recently experienced great improvements in structure resolution due to direct electron detectors with improved contrast and fast read-out leading to single electron counting. High frames rates enabled dose fractionation, where a long exposure is broken into a movie, permitting specimen drift to be registered and corrected. The typical approach for image registration, with high shot noise and low contrast, is multi-reference (MR) cross-correlation. Here we present the software package Zorro, which provides robust drift correction for dose fractionation by use of an intensity-normalized cross-correlation and logistic noise model to weight each cross-correlation in the MR model and filter each cross-correlation optimally. Frames are reliably registered by Zorro with low dose and defocus. Methods to evaluate performance are presented, by use of independently-evaluated even- and odd-frame stacks by trajectory comparison and Fourier ring correlation. Alignment of tiled sub-frames is also introduced, and demonstrated on an example dataset. Zorro source code is available at github.com/CINA/zorro. Copyright © 2016 Elsevier Inc. All rights reserved.

CryoSat Data Quality Overview

Science.gov (United States)

Bouzinac, Catherine; Fornari, Marco; Mannan, Rubinder; Parrinello, Tommaso

2013-09-01

The main objective of CryoSat mission is the observation of ice thickness with SAR interferometry. CryoSat data are routinely controlled by ESA-ESRIN- SPPA office with various tools and with the support of the IDEAS industrial consortium. This presentation introduces to the different instrumental modes and processing levels, the available quality tools (Quality Checks and Monitoring Facility) and visualisation tools, and shows the main statistical results from the data acquired and processed with the new processing version (since February 2012). A major data quality improvement is obtained from this new release of the specialized processors for all instrument modes. These new processors are also used this year for a full reprocessing campaign. This presentation also details the main improvements expected from the next release of the specialized processors in 2013.

Electron holography for fields in solids: Problems and progress

International Nuclear Information System (INIS)

Lichte, Hannes; Börrnert, Felix; Lenk, Andreas; Lubk, Axel; Röder, Falk; Sickmann, Jan; Sturm, Sebastian; Vogel, Karin; Wolf, Daniel

2013-01-01

Electron holography initially was invented by Dennis Gabor for solving the problems raised by the aberrations of electron lenses in Transmission Electron Microscopy. Nowadays, after hardware correction of aberrations allows true atomic resolution of the structure, for comprehensive understanding of solids, determination of electric and magnetic nanofields is the most challenging task. Since fields are phase objects in the TEM, electron holography is the unrivaled method of choice. After more than 40 years of experimental realization and steady improvement, holography is increasingly contributing to these highly sophisticated and essential questions in materials science, as well to the understanding of electron waves and their interaction with matter. - Highlights: • We review the development of the method of electron holography. • We outline the role of information content as guideline. • We outline the improvements of the method. • We sketch the future instrumental development. • We summarize the still existing problems to solve

Stack Characterization in CryoSat Level1b SAR/SARin Baseline C

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; Di Giacinto, Andrea; Bouffard, Jerome; Féménias, Pierre; Parrinello, Tommaso

2015-04-01

CryoSat was launched on the 8th April 2010 and is the first European ice mission dedicated to the monitoring of precise changes in the thickness of polar ice sheets and floating sea ice. CryoSat is the first altimetry mission operating in SAR mode and it carries an innovative radar altimeter called the Synthetic Aperture Interferometric Altimeter (SIRAL), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. The current CryoSat IPF (Instrument Processing Facility), Baseline B, was released in operation in February 2012. After more than 2 years of development, the release in operations of the Baseline C is expected in the first half of 2015. It is worth recalling here that the CryoSat SAR/SARin IPF1 generates 20Hz waveforms in correspondence of an approximately equally spaced set of ground locations on the Earth surface, i.e. surface samples, and that a surface sample gathers a collection of single-look echoes coming from the processed bursts during the time of visibility. Thus, for a given surface sample, the stack can be defined as the collection of all the single-look echoes pointing to the current surface sample, after applying all the necessary range corrections. The L1B product contains the power average of all the single-look echoes in the stack: the multi-looked L1B waveform. This reduces the data volume, while removing some information contained in the single looks, useful for characterizing the surface and modelling the L1B waveform. To recover such information, a set of parameters has been added to the L1B product: the stack characterization or beam behaviour parameters. The stack characterization, already included in previous Baselines, has been reviewed and expanded in Baseline C. This poster describes all the stack characterization parameters, detailing what they represent and how they have been computed. In details, such parameters can be summarized in: - Stack

ESA CryoVEx 2012

DEFF Research Database (Denmark)

Skourup, Henriette; Einarsson, Indriði; Forsberg, René

Institute (AWI) with fixed‐wing airplane (Polar‐5, Basler BT‐67). The airborne systems are described in detail, together with campaign implementation plan, data processing and data quality analysis. The CryoVEx 2012 campaign was a success and the processed data is of high quality. The data set includes 16...

Thon rings from amorphous ice and implications of beam-induced Brownian motion in single particle electron cryo-microscopy

Energy Technology Data Exchange (ETDEWEB)

McMullan, G., E-mail: gm2@mrc-lmb.cam.ac.uk; Vinothkumar, K.R.; Henderson, R.

2015-11-15

We have recorded dose-fractionated electron cryo-microscope images of thin films of pure flash-frozen amorphous ice and pre-irradiated amorphous carbon on a Falcon II direct electron detector using 300 keV electrons. We observe Thon rings [1] in both the power spectrum of the summed frames and the sum of power spectra from the individual frames. The Thon rings from amorphous carbon images are always more visible in the power spectrum of the summed frames whereas those of amorphous ice are more visible in the sum of power spectra from the individual frames. This difference indicates that while pre-irradiated carbon behaves like a solid during the exposure, amorphous ice behaves like a fluid with the individual water molecules undergoing beam-induced motion. Using the measured variation in the power spectra amplitude with number of electrons per image we deduce that water molecules are randomly displaced by a mean squared distance of ∼1.1 Å{sup 2} for every incident 300 keV e{sup −}/Å{sup 2}. The induced motion leads to an optimal exposure with 300 keV electrons of 4.0 e{sup −}/Å{sup 2} per image with which to observe Thon rings centred around the strong 3.7 Å scattering peak from amorphous ice. The beam-induced movement of the water molecules generates pseudo-Brownian motion of embedded macromolecules. The resulting blurring of single particle images contributes an additional term, on top of that from radiation damage, to the minimum achievable B-factor for macromolecular structure determination. - Highlights: • Thon rings can be seen from amorphous ice. • Radiation damage to amorphous ice randomly displaces water molecules. • Each incident 300 keV e{sup −}/Å{sup 2} displaces water molecules on average by ∼1 Å. • Macromolecules embedded in amorphous ice undergo beam induced Brownian motion.

Imaging of Norway spruce early somatic embryos with the ESEM, Cryo-SEM and laser scanning microscope

Czech Academy of Sciences Publication Activity Database

Neděla, Vilém; Hřib, Jiří; Havel, L.; Hudec, Jiří; Runštuk, Jiří

2016-01-01

Roč. 84, May 2016 (2016), s. 67-71 ISSN 0968-4328 R&D Projects: GA ČR(CZ) GA14-22777S Institutional support: RVO:68081731 Keywords : ESEM * Cryo-SEM * bright field/dark field microscopy * extracellular matrix * Picea abies * somatic embryogenesis Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.980, year: 2016

A novel storage system for cryoEM samples.

Science.gov (United States)

Scapin, Giovanna; Prosise, Winifred W; Wismer, Michael K; Strickland, Corey

2017-07-01

We present here a new CryoEM grid boxes storage system designed to simplify sample labeling, tracking and retrieval. The system is based on the crystal pucks widely used by the X-ray crystallographic community for storage and shipping of crystals. This system is suitable for any cryoEM laboratory, but especially for large facilities that will need accurate tracking of large numbers of samples coming from different sources. Copyright © 2017. Published by Elsevier Inc.

Allosteric effects in bacteriophage HK97 procapsids revealed directly from covariance analysis of cryo EM data.

Science.gov (United States)

Xu, Nan; Veesler, David; Doerschuk, Peter C; Johnson, John E

2018-05-01

The information content of cryo EM data sets exceeds that of the electron scattering potential (cryo EM) density initially derived for structure determination. Previously we demonstrated the power of data variance analysis for characterizing regions of cryo EM density that displayed functionally important variance anomalies associated with maturation cleavage events in Nudaurelia Omega Capensis Virus and the presence or absence of a maturation protease in bacteriophage HK97 procapsids. Here we extend the analysis in two ways. First, instead of imposing icosahedral symmetry on every particle in the data set during the variance analysis, we only assume that the data set as a whole has icosahedral symmetry. This change removes artifacts of high variance along icosahedral symmetry axes, but retains all of the features previously reported in the HK97 data set. Second we present a covariance analysis that reveals correlations in structural dynamics (variance) between the interior of the HK97 procapsid with the protease and regions of the exterior (not seen in the absence of the protease). The latter analysis corresponds well with hydrogen deuterium exchange studies previously published that reveal the same correlation. Copyright © 2018 Elsevier Inc. All rights reserved.

Combining image processing and modeling to generate traces of beta-strands from cryo-EM density images of beta-barrels.

Science.gov (United States)

Si, Dong; He, Jing

2014-01-01

Electron cryo-microscopy (Cryo-EM) technique produces 3-dimensional (3D) density images of proteins. When resolution of the images is not high enough to resolve the molecular details, it is challenging for image processing methods to enhance the molecular features. β-barrel is a particular structure feature that is formed by multiple β-strands in a barrel shape. There is no existing method to derive β-strands from the 3D image of a β-barrel at medium resolutions. We propose a new method, StrandRoller, to generate a small set of possible β-traces from the density images at medium resolutions of 5-10Å. StrandRoller has been tested using eleven β-barrel images simulated to 10Å resolution and one image isolated from the experimentally derived cryo-EM density image at 6.7Å resolution. StrandRoller was able to detect 81.84% of the β-strands with an overall 1.5Å 2-way distance between the detected and the observed β-traces, if the best of fifteen detections is considered. Our results suggest that it is possible to derive a small set of possible β-traces from the β-barrel cryo-EM image at medium resolutions even when no separation of the β-strands is visible in the images.

Chromosomal aberration

International Nuclear Information System (INIS)

Ishii, Yutaka

1988-01-01

Chromosomal aberrations are classified into two types, chromosome-type and chromatid-type. Chromosom-type aberrations include terminal deletion, dicentric, ring and interstitial deletion, and chromatid-type aberrations include achromatic lesion, chromatid deletion, isochromatid deletion and chromatid exchange. Clastogens which induce chromosomal aberration are divided into ''S-dependent'' agents and ''S-independent''. It might mean whether they can induce double strand breaks independent of the S phase or not. Double strand breaks may be the ultimate lesions to induce chromosomal aberrations. Caffeine added even in the G 2 phase appeared to modify the frequency of chromatid aberrations induced by X-rays and mitomycin C. Those might suggest that the G 2 phase involves in the chromatid aberration formation. The double strand breaks might be repaired by ''G 2 repair system'', the error of which might yield breakage types of chromatid aberrations and the by-pass of which might yield chromatid exchanges. Chromosome-type aberrations might be formed in the G 1 phase. (author)

An accurate optical design method for synchrotron radiation beamlines with wave-front aberration theory

Energy Technology Data Exchange (ETDEWEB)

Yu, Xiaojiang, E-mail: slsyxj@nus.edu.sg; Diao, Caozheng; Breese, Mark B. H. [Singapore Synchrotron Light Source, National University of Singapore, Singapore 117603 (Singapore)

2016-07-27

An aberration calculation method which was developed by Lu [1] can treat individual aberration term precisely. Spectral aberration is the linear sum of these aberration terms, and the aberrations of multi-element systems also can be calculated correctly when the stretching ratio, defined herein, is unity. Evaluation of focusing mirror-grating systems which are optimized according to Lu’s method, along with the Light Path Function (LPF) and the Spot Diagram method (SD) are discussed to confirm the advantage of Lu’s methodology. Lu’s aberration terms are derived from a precise wave-front treatment, whereas the terms of the power series expansion of the light path function do not yield an accurate sum of the aberrations. Moreover, Lu’s aberration terms can be individually optimized. This is not possible with the analytical spot diagram formulae.

The mechanism of DNA ejection in the Bacillus anthracis spore-binding phage 8a revealed by cryo-electron tomography

Energy Technology Data Exchange (ETDEWEB)

Fu, Xiaofeng [Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, Houston, TX 77030 (United States); Walter, Michael H. [Department of Biology, University of Northern Iowa, Cedar Falls, IA 50614 (United States); Paredes, Angel [Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, Houston, TX 77030 (United States); Morais, Marc C., E-mail: mcmorais@utmb.edu [Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX 77555 (United States); Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555 (United States); Liu, Jun, E-mail: Jun.Liu.1@uth.tmc.edu [Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, Houston, TX 77030 (United States)

2011-12-20

The structure of the Bacillus anthracis spore-binding phage 8a was determined by cryo-electron tomography. The phage capsid forms a T = 16 icosahedron attached to a contractile tail via a head-tail connector protein. The tail consists of a six-start helical sheath surrounding a central tail tube, and a structurally novel baseplate at the distal end of the tail that recognizes and attaches to host cells. The parameters of the icosahedral capsid lattice and the helical tail sheath suggest protein folds for the capsid and tail-sheath proteins, respectively, and indicate evolutionary relationships to other dsDNA viruses. Analysis of 2518 intact phage particles show four distinct conformations that likely correspond to four sequential states of the DNA ejection process during infection. Comparison of the four observed conformations suggests a mechanism for DNA ejection, including the molecular basis underlying coordination of tail sheath contraction and genome release from the capsid.

Local chromatic correction scheme for LER of PEP-II

International Nuclear Information System (INIS)

Forest, E.; Robin, D.; Zholents, A.; Donald, M.; Helm, R.; Irwin, J.; Sullivan, M.K.

1993-01-01

The correction of the chromaticity of low-beta insertions in storage rings is usually made with sextupole lenses in the ring arcs. When decreasing the beta functions at the interaction point (IP), this technique becomes fairly ineffective, since it fails to properly correct the higher-order chromatic aberrations. Here we consider the approach for ampersand PEP-II B Factory low energy ring (LER) where the chromatic effects of the quadrupole lenses generating low beta functions at the IP are corrected locally with two families of sextupoles, one family for each plane. For the IP straight section the lattice is designed in such a way that the chromatic aberrations are made small and sextupole-like aberrations are eliminated. The results of dimensional tracking simulations are presented

78 FR 49365 - Electronic Fund Transfers (Regulation E); Correction

Science.gov (United States)

2013-08-14

... BUREAU OF CONSUMER FINANCIAL PROTECTION 12 CFR Part 1005 [Docket No. CFPB-2012-0050] RIN 3170-AA33 Electronic Fund Transfers (Regulation E); Correction AGENCY: Bureau of Consumer Financial Protection. ACTION... 2013 Final Rule, which along with three other final rules \\1\\ implements the Electronic Fund Transfer...

Radiative corrections to chargino production in electron-positron collisions with polarized beams

International Nuclear Information System (INIS)

Diaz, Marco A.; King, Stephen F.; Ross, Douglas A.

2001-01-01

We study radiative corrections to chargino production at linear colliders with polarized electron beams. We calculate the one-loop corrected cross sections for polarized electon beams due to three families of quarks and squarks, working in the {ovr MS} scheme, extending our previous calculation of the unpolarized cross section with one-loop corrections due to the third family of quarks and squarks. In some cases we find rather large corrections to the tree-level cross sections. For example, for the case of right-handed polarized electrons and large tanβ the corrections can be of order 30%, allowing sensitivity to the squark mass parameters

Spectral estimation for characterization of acoustic aberration.

Science.gov (United States)

Varslot, Trond; Angelsen, Bjørn; Waag, Robert C

2004-07-01

Spectral estimation based on acoustic backscatter from a motionless stochastic medium is described for characterization of aberration in ultrasonic imaging. The underlying assumptions for the estimation are: The correlation length of the medium is short compared to the length of the transmitted acoustic pulse, an isoplanatic region of sufficient size exists around the focal point, and the backscatter can be modeled as an ergodic stochastic process. The motivation for this work is ultrasonic imaging with aberration correction. Measurements were performed using a two-dimensional array system with 80 x 80 transducer elements and an element pitch of 0.6 mm. The f number for the measurements was 1.2 and the center frequency was 3.0 MHz with a 53% bandwidth. Relative phase of aberration was extracted from estimated cross spectra using a robust least-mean-square-error method based on an orthogonal expansion of the phase differences of neighboring wave forms as a function of frequency. Estimates of cross-spectrum phase from measurements of random scattering through a tissue-mimicking aberrator have confidence bands approximately +/- 5 degrees wide. Both phase and magnitude are in good agreement with a reference characterization obtained from a point scatterer.

Conceptual Design of the Superconducting Proton Linac Short Cryo-module

CERN Document Server

Bourcey, N; Capatina, O; Azevedo, P; Montesinos, E; Parma, V; Renaglia, T; Vande Craen, A; Williams, L R; Weingarten, W; Rousselot, S; Duthil, P; Duchesne, P; Reynet, D; Dambre, P

2012-01-01

The Superconducting Proton Linac (SPL) is an R&amp;amp;D effort conducted by CERN in partnership with other international laboratories, aimed at developing key technologies for the construction of a multi-megawatt proton linac based on state-of-the-art Superconducting Radio Frequency technology, which would serve as a driver for new physics facilities such as neutrinos and radioactive ion beams. Amongst the main objectives of this effort, are the development of 704 MHz bulk niobium b=1 elliptical cavities, operating at 2 K and providing an accelerating field of 25 MV/m, and testing of a string of cavities integrated in a machine-type cryo-module. In an initial phase only four out of the eight cavities of an SPL cryo-module will be tested in a ½ length cryo-module developed for this purpose, and therefore called the Short Cryo-module. This paper presents the conceptual design of the SC, highlighting its innovative principles in terms of cavity supporting and alignment, and describes the integratio...

Electron beam position stabilization with a piezo-electric optical correction system

CERN Document Server

Averett, T; McKeown, R D; Pitt, M

1999-01-01

A piezo-electrically controlled optical correction system was successfully used to reduce the helicity-correlated pulse-to-pulse position differences of a laser spot to better than +-100 nm at a pulse rate of 600 Hz. Using a simple feedback algorithm, average position differences of DELTA x-bar=-3.5+-4.2 nm and DELTA y-bar=2.6+-6.6 nm were obtained over a 6 h period. This optical correction system was successfully used in the polarized electron source at the Bates Linear Accelerator Center to stabilize the position of the electron beam during the recent SAMPLE experiment. Because this experiment measures a parity violating signal at the 10 sup - sup 6 level, it is sensitive to systematic effects which are correlated with the helicity of the incident electrons. One potentially large systematic effect is the helicity-correlated motion of the incident electron beam. By using this optical correction system, electron beam position differences at the location of the experiment were routinely kept well below +-100 n...

Research on automatic correction of electronic beam path for distributed control

International Nuclear Information System (INIS)

Guo Xin; Su Haijun; Li Deming; Wang Shengli; Guo Honglei

2014-01-01

Background: Dynamitron, an electron irradiation accelerator of high-voltage, is used as a radiation source for industrial and agricultural production. The control system is an important component of dynamitron. Purpose: The aim is to improve the control system to meet the performance requirements of dynamitron for the stability. Methods: A distributed control system for the 1.5-MeV dynamitron is proposed to gain better performance. On this basis, an electron beam trajectory automatic correction method based on Cerebellar Model Articulation Controller and Proportional-Integral Derivative (CMAC-PID) controller is designed to improve the function of electron beam extraction system. Results: The distributed control system can meet the control requirements of the accelerator. The stability of the CMAC PID controller is better than that of conventional PID controller for the electron beam trajectory automatic correction system, and hence the CMAC-PID controller can provide better protection of dynamitron when electron beam deflection occurs. Conclusion: The distributed control system and the electron beam trajectory automatic correction method system can effectively improve the performance and reduce the failure probability of the accelerator, thereby enhancing the efficiency of the accelerator. (authors)

Mass loss of Greenland from GRACE, IceSat and CryoSat

DEFF Research Database (Denmark)

Forsberg, René; Sørensen, Louise Sandberg; Fredenslund Levinsen, Joanna

2013-01-01

CryoSat data over the Greenland Ice Sheet are used to continue elevation height change data over the Greenland ice sheet, following a decade of detailed monitoring of ice sheet changes with GRACE and IceSat. The combination and validation of the different data for measuring changes is quantified...... by using available airborne lidar data from IceBridge and CryoVEx. There is a special challenge of using CryoSat as fill-in between EnviSat and Sentinel-3 for the longterm measurements of surface elevation changes, a key essential climate variable in the ESA Climate Change Initiative. Another challenge...... is the joint utilization of both altimetry and gravity field change measurements for consistent estimates of regional change patterns. In the paper we analyze GRACE, IceSat and CryoSat data since 2003, and present consistent estimates of overall mass changes with average values around -220 GT/year, showing...

Combining Envisat and CryoSat-2 altimetry to inform hydrodynamic models

DEFF Research Database (Denmark)

Schneider, Raphael; Nygaard Godiksen, Peter; Ridler, Marc-Etienne

2016-01-01

fitted to the CryoSat-2 data: In a first step, the average simulated water levels along the river were calibrated to the CryoSat-2 data by adapting the hydrodynamic cross section datums. Subsequently the simulated water level amplitudes were fitted to those obtained from Envisat virtual station time......Remote sensing provides valuable data for parameterization and updating of hydrological models, for example water level measurements of inland water bodies from satellite radar altimeters. Many studies have used satellite altimetry data from repeat-orbit missions such as Envisat, ERS or Jason...... on Sentinel-3. CryoSat-2 SARIn level 2 data is used to improve a 1D hydrodynamic model of the Brahmaputra river basin in South Asia set up in the DHI MIKE 11 software. CryoSat-2 water levels were extracted over river masks derived from Landsat imagery. After discharge calibration, simulated water levels were...

Correction of aberrations in beams filling elliptical phase-space areas

International Nuclear Information System (INIS)

Wollnik, H.

1988-01-01

For the optimization of an optical system it is advantageous to amend the system by a virtual object lens so that the calculation always starts from an upright phase-space distribution. Furthermore, in case of a beam filling an elliptical phase-space volume, the most extreme rays of a beam, filling a parallelogram-like phase-space volume, do not exist, so that the corresponding sum of aberrations is smaller. For an optimization thus corresponding attenuation factors should be taken into accout

Improved cryo-resistors with low temperature coefficients

International Nuclear Information System (INIS)

Warnecke, P.; Braun, E.

1989-01-01

A new type of 10- and 12.9κΩ cryo-resistors operating in a liquid helium bath with small temperature coefficient of resistivity have been built. Details for the fabrication of these improved cryo-resistors are reported. Experimental evidence of their drift rates are on the order of a few parts in 10 9 per day. A reduction of the mean pressure of 98.7 kPa in the helium dewar to 86.1 kPa, corresponding to a temperature decrease from 4.19 to 4.07 Κ, did not change the resistance value by more than the experimental resolution of 4 parts in 10 8

Long helical filaments are not seen encircling cells in electron cryotomograms of rod-shaped bacteria

International Nuclear Information System (INIS)

Swulius, Matthew T.; Chen, Songye; Jane Ding, H.; Li, Zhuo; Briegel, Ariane; Pilhofer, Martin; Tocheva, Elitza I.; Lybarger, Suzanne R.; Johnson, Tanya L.; Sandkvist, Maria; Jensen, Grant J.

2011-01-01

Highlights: → No long helical filaments are seen near or along rod-shaped bacterial inner membranes by electron cryo-tomography. → Electron cryo-tomography has the resolution to detect single filaments in vivo. -- Abstract: How rod-shaped bacteria form and maintain their shape is an important question in bacterial cell biology. Results from fluorescent light microscopy have led many to believe that the actin homolog MreB and a number of other proteins form long helical filaments along the inner membrane of the cell. Here we show using electron cryotomography of six different rod-shaped bacterial species, at macromolecular resolution, that no long (>80 nm) helical filaments exist near or along either surface of the inner membrane. We also use correlated cryo-fluorescent light microscopy (cryo-fLM) and electron cryo-tomography (ECT) to identify cytoplasmic bundles of MreB, showing that MreB filaments are detectable by ECT. In light of these results, the structure and function of MreB must be reconsidered: instead of acting as a large, rigid scaffold that localizes cell-wall synthetic machinery, moving MreB complexes may apply tension to growing peptidoglycan strands to ensure their orderly, linear insertion.

Advanced Electron Microscopy in Materials Physics

International Nuclear Information System (INIS)

Zhu, Y.; Jarausch, K.

2009-01-01

Aberration correction has opened a new frontier in electron microscopy by overcoming the limitations of conventional round lenses, providing sub-angstrom-sized probes and extending information limits. The imaging and analytical performance of these corrector-equipped microscopes affords an unprecedented opportunity to study structure-property relationships of matter at the atomic scale. This new generation of microscopes is able to retrieve high-quality structural information comparable to neutron and synchrotron x-ray experiments, but with local atomic resolution. These advances in instrumentation are accelerating the research and development of various functional materials ranging from those for energy generation, conversion, transportation and storage to those for catalysis and nano-device applications. The dramatic improvements in electron-beam illumination and detection also present a host of new challenges for the interpretation and optimization of experiments. During 7-9 November 2007, a workshop, entitled 'Aberration Corrected Electron Microscopy in Material Physics', was convened at the Center for Functional Nanomaterials, Brookhaven National Laboratories (BNL) to address these opportunities and challenges. The workshop was co-sponsored by Hitachi High Technologies, a leader in electron microscopy instrumentation, and BNL's Institute of Advanced Electron Microscopy, a leader in materials physics research using electron microscopy. The workshop featured presentations by internationally prominent scientists working at the frontiers of electron microscopy, both on developing instrumentation and applying it in materials physics. The meeting, structured to stimulate scientific exchanges and explore new capabilities, brought together ∼100 people from over 10 countries. This special issue complies many of the advances in instrument performance and materials physics reported by the invited speakers and attendees at the workshop.

Transport and installation of the LHC cryo-magnets

CERN Document Server

Artoos, K; Capatina, O; Chevalley, JM; Foraz, K; Guinchard, M; Hauviller, Claude; Kershaw, K; Prodon, S; Rühl, Ingo; Trinquart, G; Weisz, S; Ponsot, P

2007-01-01

Eleven years have passed between the beginning of transport and handling studies in 1996 and the completion of the LHC cryo-magnets installation in 2007. More than 1700 heavy, long and fragile cryo-magnets had to be transported and installed in the 27 km long LHC tunnel with very restricted available space. The size and complexity of the project involved challenges in the field of equipment design and manufacturing, maintenance, training and follow-up of operators and logistics. The paper presents the milestones, problems to be overcome and lessons learned during this project.

A model of distributed phase aberration for deblurring phase estimated from scattering.

Science.gov (United States)

Tillett, Jason C; Astheimer, Jeffrey P; Waag, Robert C

2010-01-01

Correction of aberration in ultrasound imaging uses the response of a point reflector or its equivalent to characterize the aberration. Because a point reflector is usually unavailable, its equivalent is obtained using statistical methods, such as processing reflections from multiple focal regions in a random medium. However, the validity of methods that use reflections from multiple points is limited to isoplanatic patches for which the aberration is essentially the same. In this study, aberration is modeled by an offset phase screen to relax the isoplanatic restriction. Methods are developed to determine the depth and phase of the screen and to use the model for compensation of aberration as the beam is steered. Use of the model to enhance the performance of the noted statistical estimation procedure is also described. Experimental results obtained with tissue-mimicking phantoms that implement different models and produce different amounts of aberration are presented to show the efficacy of these methods. The improvement in b-scan resolution realized with the model is illustrated. The results show that the isoplanatic patch assumption for estimation of aberration can be relaxed and that propagation-path characteristics and aberration estimation are closely related.

Using Aberrant Behaviors as Reinforcers for Autistic Children.

Science.gov (United States)

Charlop, Marjorie H.; And Others

1990-01-01

Three experiments assessed the efficacy of various reinforcers to increase correct task responding in a total of 10 autistic children, aged 6-9. Of the reinforcers used (stereotypy, delayed echolalia, perseverative behavior, and food), task performance was highest with opportunities to engage in aberrant behaviors, and lowest with edible…

Experimental investigations of chromosomal aberrations following irradiation with fast electrons at various irradiation depths as influenced by sulfoguanidine

International Nuclear Information System (INIS)

Donnerstag, R.

1975-01-01

Root tips of Vicia faba were irradiated with fast electrons (14.2 MeV, 120 rad) in the phantom at a depth of 100% and 30% relative depth dose in culture medium and sulphoguanidine solution and were fixed after 3, 6, 9, 12 and 24 h. In contrast to the controls, a transitory mitotic depression was observed after irradiation. This effect was more marked for root tips irradiated in culture medium and much less pronounced in root tips irradiated in sulphoguanidine. When quantitatively assessing the anaphasal aberrations, the highest damage rate was found at 3 h p.r. This was attributed to the facts that these cells had been irradiated in the earliest G 2 phase. A second aberration peak was found 9 h after irradiation at 30% relative depth dose. These cells had been irradiated in the middle of the S phase. It is assumed that with increasing LET, euchromatic DNA regions are most readily damaged. When the two irradiation depths were compared, it was found that the aberration rate was significantly reduced after irradiation in sulphoguanidine solution at a depth of 30% relative dose. A qualitative assessment showed that fragments made up the highest and bridges the lowest fraction of aberrations, although there were variations in the single values depending on the irradiation conditions and culture media used. It was assumed that sulfoguanidine may have a positive influence on repair processes in damaged DNA, and that this influence depends on the energy spectrum of the irradiation. Furthermore, the possibility of a biochemical mechanism was discussed. (orig./MG) [de

Theory of aberration fields for general optical systems with freeform surfaces.

Science.gov (United States)

Fuerschbach, Kyle; Rolland, Jannick P; Thompson, Kevin P

2014-11-03

This paper utilizes the framework of nodal aberration theory to describe the aberration field behavior that emerges in optical systems with freeform optical surfaces, particularly φ-polynomial surfaces, including Zernike polynomial surfaces, that lie anywhere in the optical system. If the freeform surface is located at the stop or pupil, the net aberration contribution of the freeform surface is field constant. As the freeform optical surface is displaced longitudinally away from the stop or pupil of the optical system, the net aberration contribution becomes field dependent. It is demonstrated that there are no new aberration types when describing the aberration fields that arise with the introduction of freeform optical surfaces. Significantly it is shown that the aberration fields that emerge with the inclusion of freeform surfaces in an optical system are exactly those that have been described by nodal aberration theory for tilted and decentered optical systems. The key contribution here lies in establishing the field dependence and nodal behavior of each freeform term that is essential knowledge for effective application to optical system design. With this development, the nodes that are distributed throughout the field of view for each aberration type can be anticipated and targeted during optimization for the correction or control of the aberrations in an optical system with freeform surfaces. This work does not place any symmetry constraints on the optical system, which could be packaged in a fully three dimensional geometry, without fold mirrors.

Brown's TRANSPORT up to third order aberration by artificial intelligence

International Nuclear Information System (INIS)

Xia Jiawen; Xie Xi; Qiao Qingwen

1991-01-01

Brown's TRANSPORT is a first and second order matrix multiplication computer program intended for the design of accelerator beam transport systems, neglecting the third order aberration. Recently a new method was developed to derive analytically any order aberration coefficients of general charged particle optic system, applicable to any practical systems, such as accelerators, electron microscopes, lithographs, etc., including those unknown systems yet to be invented. An artificial intelligence program in Turbo Prolog was implemented on IBM-PC 286 or 386 machine to generate automatically the analytical expression of any order aberration coefficients of general charged particle optic system. Based on this new method and technique, Brown's TRANSPORT is extended beyond the second order aberration effects by artificial intelligence, outputing automatically all the analytical expressions up to the third order aberration coefficients

Brown's transport up to third order aberration by artificial intelligence

International Nuclear Information System (INIS)

Xia Jiawen; Xie Xi; Qiao Qingwen

1992-01-01

Brown's TRANSPORT is a first and second order matrix multiplication computer program intended for the design of accelerator beam transport systems, neglecting the third order aberration. Recently a new method was developed to derive analytically any order aberration coefficients of general charged particle optic system, applicable to any practical systems, such as accelerators, electron microscopes, lithographs, including those unknown systems yet to be invented. An artificial intelligence program in Turbo Prolog was implemented on IBM-PC 286 or 386 machine to generate automatically the analytical expression of any order aberration coefficients of general charged particle optic system. Based on this new method and technique, Brown's TRANSPORT is extended beyond the second order aberration effect by artificial intelligence, outputting automatically all the analytical expressions up to the third order aberration coefficients

Static telescope aberration measurement using lucky imaging techniques

Science.gov (United States)

López-Marrero, Marcos; Rodríguez-Ramos, Luis Fernando; Marichal-Hernández, José Gil; Rodríguez-Ramos, José Manuel

2012-07-01

A procedure has been developed to compute static aberrations once the telescope PSF has been measured with the lucky imaging technique, using a nearby star close to the object of interest as the point source to probe the optical system. This PSF is iteratively turned into a phase map at the pupil using the Gerchberg-Saxton algorithm and then converted to the appropriate actuation information for a deformable mirror having low actuator number but large stroke capability. The main advantage of this procedure is related with the capability of correcting static aberration at the specific pointing direction and without the need of a wavefront sensor.

Propagation of highly aberrated laser beams in nonquadratic plasma waveguides

International Nuclear Information System (INIS)

Feit, M.D.; Fleck, J.A. Jr.; Morris, J.R.

1977-01-01

The propagation of a laser beam in a plasma column several meters long with a realistic electron density distribution is examined. The electron density distribution is based on laser-beam heating at z=0, but is otherwise uncoupled to the laser beam. The aberrated nature of the resulting lenslike medium leads to essentially aperiodic beam properties, which contrast with the completely periodic properties of Gaussian beams propagating in quadratic lenslike media. The beam is nonetheless stably trapped. These aberrated-beam properties also help to stabilize the beam against axial variations in refractive index

Precision controlled atomic resolution scanning transmission electron microscopy using spiral scan pathways

Science.gov (United States)

Sang, Xiahan; Lupini, Andrew R.; Ding, Jilai; Kalinin, Sergei V.; Jesse, Stephen; Unocic, Raymond R.

2017-03-01

Atomic-resolution imaging in an aberration-corrected scanning transmission electron microscope (STEM) can enable direct correlation between atomic structure and materials functionality. The fast and precise control of the STEM probe is, however, challenging because the true beam location deviates from the assigned location depending on the properties of the deflectors. To reduce these deviations, i.e. image distortions, we use spiral scanning paths, allowing precise control of a sub-Å sized electron probe within an aberration-corrected STEM. Although spiral scanning avoids the sudden changes in the beam location (fly-back distortion) present in conventional raster scans, it is not distortion-free. “Archimedean” spirals, with a constant angular frequency within each scan, are used to determine the characteristic response at different frequencies. We then show that such characteristic functions can be used to correct image distortions present in more complicated constant linear velocity spirals, where the frequency varies within each scan. Through the combined application of constant linear velocity scanning and beam path corrections, spiral scan images are shown to exhibit less scan distortion than conventional raster scan images. The methodology presented here will be useful for in situ STEM imaging at higher temporal resolution and for imaging beam sensitive materials.

The effect of quantum correction on plasma electron heating in ultraviolet laser interaction

Energy Technology Data Exchange (ETDEWEB)

Zare, S.; Sadighi-Bonabi, R., E-mail: Sadighi@sharif.ir; Anvari, A. [Department of Physics, Sharif University of Technology, P.O. Box 11365-9567, Tehran (Iran, Islamic Republic of); Yazdani, E. [Department of Energy Engineering and Physics, Amirkabir University of Technology, P.O. Box 15875-4413, Tehran (Iran, Islamic Republic of); Hora, H. [Department of Theoretical Physics, University of New South Wales, Sydney 2052 (Australia)

2015-04-14

The interaction of the sub-picosecond UV laser in sub-relativistic intensities with deuterium is investigated. At high plasma temperatures, based on the quantum correction in the collision frequency, the electron heating and the ion block generation in plasma are studied. It is found that due to the quantum correction, the electron heating increases considerably and the electron temperature uniformly reaches up to the maximum value of 4.91 × 10{sup 7 }K. Considering the quantum correction, the electron temperature at the laser initial coupling stage is improved more than 66.55% of the amount achieved in the classical model. As a consequence, by the modified collision frequency, the ion block is accelerated quicker with higher maximum velocity in comparison with the one by the classical collision frequency. This study proves the necessity of considering a quantum mechanical correction in the collision frequency at high plasma temperatures.

Ten years of cryo-magnetic W7-X test facility construction and operation

International Nuclear Information System (INIS)

Renard, B.; Dispau, G.; Donati, A.; Genini, L.; Gournay, J.F.; Kuster, O.; Molinie, F.; Schild, T.; Touzery, R.; Vieillard, L.; Walter, C.

2011-01-01

The construction, commissioning, and operation phases of the W7-X cryo-magnetic test facility in CEA Saclay lasted ten years. The large diversity of equipments called, specialties involved and problems solved attest the expertise that was required to operate the test facility and test the coils. Nearly one hundred cryogenic tests were performed on the seventy W7-X coils, at a rate always increasing, using two cryostats each holding two coils. This paper presents the test facility and its operation first, the cryogenic difficulties that were confronted with their solutions, the electro-magnetic difficulties encountered along with corrective actions, and finally the instrumentation and data acquisition aspects. (authors)

Correlative Cryo-Tem Cryo-Stxm and Cryo-Shxm Investigation of Selenium Bioreduction in a Contaminated Aquifer

Science.gov (United States)

Fakra, S.; Luef, B.; Tyliszczak, T.; Castelle, C. J.; Mullin, S. W.; Hug, L. A.; Williams, K. H.; Marcus, M.; Banfield, J. F.

2015-12-01

Accurate mapping of the composition and ultrastructure of minerals and cells is key to understanding biogeochemical process in contaminated environments. Here we developed two apparatus that allow correlation of cryogenic transmission electron microscopy (TEM), synchrotron hard X-ray microprobe (SHXM) and scanning transmission X-ray microscopy (STXM) datasets. These cryogenic methods enabled precise determination of the distribution, valence state and structure of selenium in intact biofilms sampled during a biostimulation experiment in a contaminated aquifer near Rifle, CO, USA. Results were replicated in the laboratory via anaerobic selenate-reducing enrichment cultures. 16S rRNA analyses of field-derived biofilm indicated the dominance of Betaproteobacteria from the Comamonadaceae family, and uncultivated members of the Simplicispira genus. The major product in field and culture-derived biofilms consists of ~25-300 nm red amorphous Se0 aggregates of colloidal nanoparticles. Correlative analyses of the cultures provided direct evidence for microbial dissimilatory reduction of Se(VI) to Se(IV) to Se0. X-ray diffraction and Se K-edge extended X-ray absorption fine structure spectroscopy revealed red amorphous Se0 with a first shell Se-Se interatomic distance of 2.339 ± 0.003 Å. STXM showed that these aggregates are strongly associated with a protein-rich biofilm matrix containing acidic polysaccharides. From Rifle groundwater, we isolated a strain that shares 98.9% 16S rRNA gene sequence identity with Dechloromonas aromatica RCB and grows anaerobically by oxidizing acetate and reducing selenate. We refer to this isolate as Dechloromonas selenatis strain RGW99. 3D cryo-electron tomography showed that Se0 particles do not form inside the cytoplasm but rather originate in the cell membrane. The end product of selenate reduction by D. selenatis is 240 ± 66 nm diameter red amorphous Se0 colloidal aggregates. This product was found to be stable for months. Overall

Quantitative atom column position analysis at the incommensurate interfaces of a (PbS)1.14NbS2 misfit layered compound with aberration-corrected HRTEM

International Nuclear Information System (INIS)

Garbrecht, M.; Spiecker, E.; Tillmann, K.; Jaeger, W.

2011-01-01

Aberration-corrected HRTEM is applied to explore the potential of NCSI contrast imaging to quantitatively analyse the complex atomic structure of misfit layered compounds and their incommensurate interfaces. Using the (PbS) 1.14 NbS 2 misfit layered compound as a model system it is shown that atom column position analyses at the incommensurate interfaces can be performed with precisions reaching a statistical accuracy of ±6 pm. The procedure adopted for these studies compares experimental images taken from compound regions free of defects and interface modulations with a structure model derived from XRD experiments and with multi-slice image simulations for the corresponding NCSI contrast conditions used. The high precision achievable in such experiments is confirmed by a detailed quantitative analysis of the atom column positions at the incommensurate interfaces, proving a tetragonal distortion of the monochalcogenide sublattice. -- Research Highlights: → Quantitative aberration-corrected HRTEM analysis of atomic column positions in (PbS) 1.14 NbS 2 misfit layered compound reveals tetragonal distortion of the PbS subsystem. → Detailed comparison of multi-slice simulations with the experimental NCSI contrast condition imaging results lead to a high precision (better than 10 pm) for determining the positions of atoms. → Precision in gaining information of local structure at atomic scale is demonstrated, which may not be accessible by means of X-ray and neutron diffraction analysis.

Characterisation of colloidal drug delivery systems from the naked eye to Cryo-FESEM

DEFF Research Database (Denmark)

Krauel, Karen; Girvan, Liz; Hook, Sarah

2007-01-01

Poly(ethylcyanoacrylate) nanoparticles prepared by interfacial polymerisation on the basis of microemulsions were prepared in this study and both colloidal systems, nanoparticles and microemulsions, were analysed by visual observation and several microscopic techniques. Phase boundaries...... for the microemulsions of the two pseudoternary systems ethyloleate, polyoxyethylene 20 sorbitan mono-oleate/sorbitan monolaurate and water with and without butanol as a cosurfactant were determined by visual observation of the samples. Microemulsions containing liquid crystals were determined by polarisation light...... microscopy. Using freeze-fracture transmission electron microscopy and Cryo-field emission scanning electron microscopy the type of microemulsion (w/o droplet, bicontinuous, solution) was characterised. Nanoparticles prepared from the different types of microemulsion were additionally observed...

Imaging properties and energy aberrations of a double-pass cylindrical-mirror electron energy analyzer

International Nuclear Information System (INIS)

Erickson, N.E.; Powell, C.J.

1986-01-01

The imaging properties and energy aberrations of a commercial double-pass cylindrical-mirror analyzer have been characterized using an extension of the method recently reported by Seah and Mathieu. The electron beam from the coaxial electron gun was rastered across a test surface and the intensity of either elastically scattered electrons or of electrons at other selected energies was stored in a computer as a function of beam position on the specimen and other experimental parameters. The intensity data were later plotted to provide an ''image'' of the detected intensity. Images of this type are presented for electron energies of 100, 500, and 1000 eV and for the application of small offset voltages (typically between -1 and +5 V) between the analyzer and the gun cathode with the instrument operated in conditions appropriate for XPS or AES. Small offset voltages ( or approx. =5 V) lead to image shapes similar to those for the elastic peak but with 20%--40% increased widths. Deflection of the incident beam by up to 2 mm from the axis caused variations of up to +-0.15 eV in the measured positions of the elastic peak. Our observations can be interpreted qualitatively in terms of the known relationship between detected signal and combinations of position of electron emission from the specimen, angle of emission, and electron energy. The images obtained with elastically and inelastically scattered electrons provide a convenient and quantitative means of assessing instrument performance and of defining the specimen area being analyzed for the particular combination of instrument operating conditions and the energy width of AES or XPS features from the specimen

Preventive and corrective maintenance for MINT EPS 3000 electron beam machine

International Nuclear Information System (INIS)

Siti Aiasah Hashim; Shaari Jahar

2005-01-01

Preventive and corrective maintenance of a high energy electron is to ensure that the machine would not fail during operation. MINT's EPS 3000 electron beam machine has been in operation for almost 12 years. Throughout those years, events relating to scheduled overhauls and unscheduled corrective maintenance had provided invaluable experience in the form of informal maintenance training to the operators. With the implementation of ISO 9001:2000 Quality Management System at the facility, the preventive and corrective maintenance program is becoming more structured and orderly. Collected maintenance data shall be used to initiate continual improvement activities in the facility.This paper describes MINT-ALURTRON's 12 years experience in providing maintenance work for the EPS 3000. (Author)

Cryo-EM structure of isomeric molluscan hemocyanin triggered by viral infection.

Directory of Open Access Journals (Sweden)

Hongtao Zhu

Full Text Available Hemocyanins (Hcs of arthropods and mollusks function not only as oxygen transporters, but also as phenoloxidases (POs. In invertebrates, PO is an important component in the innate immune cascade, where it functions as the initiator of melanin synthesis, a pigment involved in encapsulating and killing of pathogenic microbes. Although structures of Hc from several species of invertebrates have been reported, the structural basis for how PO activity is triggered by structural changes of Hc in vivo remains poorly understood. Here, we report a 6.8 Å cryo-electron microscopy (cryo-EM structure of the isomeric form of hemocyanin, which was isolated from Abalone Shriveling syndrome-associated Virus (AbSV infected abalone (Halitotis diversicolor, and build a pseudoatomic model of isomeric H. diversicolor hemocyanin 1 (HdH1. Our results show that, compared with native form of HdH1, the architecture of isomeric HdH1 turns into a more relaxed form. The interactions between certain functional units (FUs present in the native form of Hc either decreased or were totally abolished in the isomeric form of Hc. As a result of that, native state Hc switches to its isomeric form, enabling it to play its role in innate immune responses against invading pathogens.

76 FR 709 - Electronic Funds Transfer of Depository Taxes; Correction

Science.gov (United States)

2011-01-06

... DEPARTMENT OF THE TREASURY Internal Revenue Service 26 CFR Parts 40 and 301 [TD 9507] RIN 1545-BJ13 Electronic Funds Transfer of Depository Taxes; Correction AGENCY: Internal Revenue Service (IRS...) providing guidance relating to Federal tax deposits (FTDs) by Electronic Funds Transfer (EFT). The temporary...

76 FR 708 - Electronic Funds Transfer of Depository Taxes; Correction

Science.gov (United States)

2011-01-06

... DEPARTMENT OF THE TREASURY Internal Revenue Service 26 CFR Parts 1, 31, 40, and 301 [TD 9507] RIN 1545-BJ13 Electronic Funds Transfer of Depository Taxes; Correction AGENCY: Internal Revenue Service... Electronic Funds Transfer (EFT). The temporary and final regulations provide rules under which depositors...

Optical Aberrations and Wavefront

Directory of Open Access Journals (Sweden)

Nihat Polat

2014-08-01

Full Text Available The deviation of light to create normal retinal image in the optical system is called aberration. Aberrations are divided two subgroup: low-order aberrations (defocus: spherical and cylindrical refractive errors and high-order aberrations (coma, spherical, trefoil, tetrafoil, quadrifoil, pentafoil, secondary astigmatism. Aberrations increase with aging. Spherical aberrations are compensated by positive corneal and negative lenticular spherical aberrations in youth. Total aberrations are elevated by positive corneal and positive lenticular spherical aberrations in elderly. In this study, we aimed to analyze the basic terms regarding optic aberrations which have gained significance recently. (Turk J Ophthalmol 2014; 44: 306-11

Pseudo LRM waveforms from CryoSat SARin acquisition

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; Bouffard, Jerome; Parrinello, Tommaso; Féménias, Pierre

2016-04-01

CryoSat was launched on the 8th April 2010 and is the first European ice mission dedicated to the monitoring of precise changes in the thickness of polar ice sheets and floating sea ice. The main payload of CryoSat is a Ku-band pulsewidth limited radar altimeter, called SIRAL (Synthetic interferometric radar altimeter). When commanded in SARIn (synthetic aperture radar interferometry) mode, through coherent along-track processing of the returns received from two antennas, the interferometric phase related to the first arrival of the echo is used to retrieve the angle of arrival of the scattering in the across-track direction. When SIRAL operates in SAR or SARin mode, the obtained waveforms have an along-track resolution and a speckle reduction which is increased with respect to the pulse-limited waveforms. Anyway, in order to analyze the continuity of the geophysical retrieved parameters among different acquisition modes, techniques to transform SARin mode data to pseudo-LRM mode data are welcome. The transformation process is known as SAR reduction and it is worth recalling here that only approximate pseudo-LRM waveforms can be obtained in case of closed burst acquisitions, as SIRAL operates. A SAR reduction processing scheme has been developed to obtain pseudo-LRM waveforms from CryoSat SARin acquisition. As a trade-off between the along-track length on Earth surface contributing to one SARin pseudo-LRM waveform and the noisiness of the waveform itself, it has been chosen a SAR reduction approach based on the averaging of all the SARin echoes received each 20Hz, resulting in one pseudo-LRM waveform for each SARin burst given the SARin burst repetition period. SARin pseudo-LRM waveforms have been produced for CryoSat acquisition both on ice and sea surfaces, aiming at verifying the continuity of the retracked surface height over the ellipsoid between genuine LRM products and pseudo-LRM products. Moreover, the retracked height from the SARin pseudo-LRM has been

3D printing of microlenses for aberration correction in GRIN microendoscopes

KAUST Repository

Antonini, Andrea; Bovetti, Serena; Moretti, Claudio; Succol, Francesca; Rajamanickam, Vijayakumar Palanisamy; Bertoncini, Andrea; Fellin, Tommaso; Liberale, Carlo

2017-01-01

Two-photon fluorescence microscopy provides high resolution information on the anatomy and function of cellular structures located several hundreds of microns deep within biological tissues. However, light scattering poses a fundamental limit to imaging of deeper areas (> 1.5 mm). Implantable microendoscopic probes based on graded index (GRIN) lenses are widely used tools to perform two-photon fluorescence microscopy in otherwise inaccessible regions[1], but the optical performances of with these probes are limited by intrinsic aberrations.

3D printing of microlenses for aberration correction in GRIN microendoscopes

KAUST Repository

Antonini, Andrea

2017-11-02

Two-photon fluorescence microscopy provides high resolution information on the anatomy and function of cellular structures located several hundreds of microns deep within biological tissues. However, light scattering poses a fundamental limit to imaging of deeper areas (> 1.5 mm). Implantable microendoscopic probes based on graded index (GRIN) lenses are widely used tools to perform two-photon fluorescence microscopy in otherwise inaccessible regions[1], but the optical performances of with these probes are limited by intrinsic aberrations.

Structure and assembly of the mouse ASC inflammasome by combined NMR spectroscopy and cryo-electron microscopy

Science.gov (United States)

Sborgi, Lorenzo; Ravotti, Francesco; Dandey, Venkata P.; Dick, Mathias S.; Mazur, Adam; Reckel, Sina; Chami, Mohamed; Scherer, Sebastian; Huber, Matthias; Böckmann, Anja; Egelman, Edward H.; Stahlberg, Henning; Broz, Petr; Meier, Beat H.; Hiller, Sebastian

2015-01-01

Inflammasomes are multiprotein complexes that control the innate immune response by activating caspase-1, thus promoting the secretion of cytokines in response to invading pathogens and endogenous triggers. Assembly of inflammasomes is induced by activation of a receptor protein. Many inflammasome receptors require the adapter protein ASC [apoptosis-associated speck-like protein containing a caspase-recruitment domain (CARD)], which consists of two domains, the N-terminal pyrin domain (PYD) and the C-terminal CARD. Upon activation, ASC forms large oligomeric filaments, which facilitate procaspase-1 recruitment. Here, we characterize the structure and filament formation of mouse ASC in vitro at atomic resolution. Information from cryo-electron microscopy and solid-state NMR spectroscopy is combined in a single structure calculation to obtain the atomic-resolution structure of the ASC filament. Perturbations of NMR resonances upon filament formation monitor the specific binding interfaces of ASC-PYD association. Importantly, NMR experiments show the rigidity of the PYD forming the core of the filament as well as the high mobility of the CARD relative to this core. The findings are validated by structure-based mutagenesis experiments in cultured macrophages. The 3D structure of the mouse ASC-PYD filament is highly similar to the recently determined human ASC-PYD filament, suggesting evolutionary conservation of ASC-dependent inflammasome mechanisms. PMID:26464513

Transmission electron microscopy of polyhydroxybutyrate-co-valerate (PHBV)/nanocrystalline cellulose (NCC) bio-nanocomposite prepared using cryo-ultramicrotomy

Science.gov (United States)

Ismarul, N. I.; Engku, A. H. E. U.; Siti, N. K.; Tay, K. Y.

2017-12-01

Environmental issues on disposal and end-of-life for product made from synthetic petroleum-derived polymers have gained increasing attention from materials scientist to search for new materials with similar physical and mechanical properties but environmental friendly in a way that they are renewable and biodegradable as well. This work is to study the effect of nanocrystalline cellulose in improving the thermal stability of polyhydroxybutyrate-co-valerate biopolymer for high temperature processing of packaging material. 10 % w/w PHBV-NCC bio-nanocomposite feedstock pellet prepared using RONDOL minilab compounder was used as the sample for the preparation of Transmission Electron Microscopy (TEM) sample. RMC Cryo-Ultramicrotomy equipment was used to prepare the ultra-thin slice of the bio-nanocomposite pellet under liquid nitrogen at - 60 °C. Diamond knife was used to slice off about 80-100 nm ultra-thin bio-nanocomposite films and was transferred into the lacey carbon film coated grid using cooled sugar solution. A few drops of phosphotungstic acid was used as negative stain to improve the contrast during the TEM analysis. HITACHI TEM systems was used to obtain the TEM micrograph of PHBV-NCC bio-nanocomposite using 80kV accelerating voltage. A well dispersed NCC in PHBV matrix, ranging from 5 to 25 nm in width was observed.

Cryo-EM structures of the human endolysosomal TRPML3 channel in three distinct states.

Science.gov (United States)

Zhou, Xiaoyuan; Li, Minghui; Su, Deyuan; Jia, Qi; Li, Huan; Li, Xueming; Yang, Jian

2017-12-01

TRPML3 channels are mainly localized to endolysosomes and play a critical role in the endocytic pathway. Their dysfunction causes deafness and pigmentation defects in mice. TRPML3 activity is inhibited by low endolysosomal pH. Here we present cryo-electron microscopy (cryo-EM) structures of human TRPML3 in the closed, agonist-activated, and low-pH-inhibited states, with resolutions of 4.06, 3.62, and 4.65 Å, respectively. The agonist ML-SA1 lodges between S5 and S6 and opens an S6 gate. A polycystin-mucolipin domain (PMD) forms a luminal cap. S1 extends into this cap, forming a 'gating rod' that connects directly to a luminal pore loop, which undergoes dramatic conformational changes in response to low pH. S2 extends intracellularly and interacts with several intracellular regions to form a 'gating knob'. These unique structural features, combined with the results of electrophysiological studies, indicate a new mechanism by which luminal pH and other physiological modulators such as PIP 2 regulate TRPML3 by changing S1 and S2 conformations.

A Validation Dataset for CryoSat Sea Ice Investigators

DEFF Research Database (Denmark)

Julia, Gaudelli,; Baker, Steve; Haas, Christian

Since its launch in April 2010 Cryosat has been collecting valuable sea ice data over the Arctic region. Over the same period ESA’s CryoVEx and NASA IceBridge validation campaigns have been collecting a unique set of coincident airborne measurements in the Arctic. The CryoVal-SI project has...... community. In this talk we will describe the composition of the validation dataset, summarising how it was processed and how to understand the content and format of the data. We will also explain how to access the data and the supporting documentation....

Performance analysis of a large-scale helium Brayton cryo-refrigerator with static gas bearing turboexpander

International Nuclear Information System (INIS)

Zhang, Yu; Li, Qiang; Wu, Jihao; Li, Qing; Lu, Wenhai; Xiong, Lianyou; Liu, Liqiang; Xu, Xiangdong; Sun, Lijia; Sun, Yu; Xie, Xiujuan; Wang, Bingming; Qiu, Yinan; Zhang, Peng

2015-01-01

Highlights: • A 2 kW at 20.0 K helium Brayton cryo-refrigerator is built in China. • A series of tests have been systematically conducted to investigate the performance of the cryo-refrigerator. • Maximum heat conductance proportion (90.7%) appears in the heat exchangers of cold box rather than those of heat reservoirs. • A model of helium Brayton cryo-refrigerator/cycle is presented according to finite-time thermodynamics. - Abstract: Large-scale helium cryo-refrigerator is widely used in superconducting systems, nuclear fusion engineering, and scientific researches, etc., however, its energy efficiency is quite low. First, a 2 kW at 20.0 K helium Brayton cryo-refrigerator is built, and a series of tests have been systematically conducted to investigate the performance of the cryo-refrigerator. It is found that maximum heat conductance proportion (90.7%) appears in the heat exchangers of cold box rather than those of heat reservoirs, which is the main characteristic of the helium Brayton cryo-refrigerator/cycle different from the air Brayton refrigerator/cycle. Other three characteristics also lie in the configuration of refrigerant helium bypass, internal purifier and non-linearity of specific heat of helium. Second, a model of helium Brayton cryo-refrigerator/cycle is presented according to finite-time thermodynamics. The assumption named internal purification temperature depth (PTD) is introduced, and the heat capacity rate of whole cycle is divided into three different regions in accordance with the PTD: room temperature region, upper internal purification temperature region and lower one. Analytical expressions of cooling capacity and COP are obtained, and we found that the expressions are piecewise functions. Further, comparison between the model and the experimental results for cooling capacity of the helium cryo-refrigerator shows that error is less than 7.6%. The PTD not only helps to achieve the analytical formulae and indicates the working

An Image Processing Approach to Pre-compensation for Higher-Order Aberrations in the Eye

Directory of Open Access Journals (Sweden)

Miguel Alonso Jr

2004-06-01

Full Text Available Human beings rely heavily on vision for almost all of the tasks that are required in daily life. Because of this dependence on vision, humans with visual limitations, caused by genetic inheritance, disease, or age, will have difficulty in completing many of the tasks required of them. Some individuals with severe visual impairments, known as high-order aberrations, may have difficulty in interacting with computers, even when using a traditional means of visual correction (e.g., spectacles, contact lenses. This is, in part, because these correction mechanisms can only compensate for the most regular (low-order distortions or aberrations of the image in the eye. This paper presents an image processing approach that will pre-compensate the images displayed on the computer screen, so as to counter the effect of the eye's aberrations on the image. The characterization of the eye required to perform this customized pre-compensation is the eye's Point Spread Function (PSF. Ophthalmic instruments generically called "Wavefront Analyzers" can now measure this description of the eye's optical properties. The characterization provided by these instruments also includes the "higher-order aberration components" and could, therefore, lead to a more comprehensive vision correction than traditional mechanisms. This paper explains the theoretical foundation of the methods proposed and illustrates them with experiments involving the emulation of a known and constant PSF by interposing a lens in the field of view of normally sighted test subjects.

Simulação computacional de cirurgias foto-refrativas personalizadas e precisão relacionada à ordem das aberrações ópticas Computational simulation of customized photorefractive surgery and precision of correction related to different order aberrations

Directory of Open Access Journals (Sweden)

Luis Alberto Vieira de Carvalho

2007-12-01

Full Text Available O objetivo principal deste trabalho foi o desenvolvimento e a implementação de um algoritmo para simulação de cirurgias foto-refrativas. Sabe-se que os atuais " flying-spot" lasers para cirurgia refrativa disponíveis no mercado ainda contêm limitações na correção de algumas aberrações de ordem alta. No entanto, há muito pouca informação quantitativa sobre os erros envolvidos em função da complexidade das aberrações. Aplicando-se superfícies objetivo descritas por uma série de coeficientes de Zernike e o conceito matemático de convolução, várias ablações personalizadas foram simuladas para várias aberrações, desde baixa ordem (esfero-cilíndricas até aberrações de décima ordem (como coma, aberração esférica, entre outras. Os resultados mostram que o perfil de ablação de cada pulso do laser assim como seu diâmetro são fatores críticos quando o objetivo é corrigir aberrações de alta ordem. Além disso, outras conclusões foram possíveis, como por exemplo a de que nem todas as aberrações de alta ordem induzem aos mesmos erros na ablação. Também foi possível verificar que a correção de aberrações de alta ordem nos termos radiais (como aberração esférica é bem mais difícil do que naqueles termos contendo alta frequência angular (como trefoil.PURPOSE: To develop and implement an algorithm for simulation of photorefractive surgery. It is well known that many flying-spot lasers in the market have limitations in correcting higher order aberrations, but there is little quantitative information about errors related to aberration complexity. METHODS: By applying known surfaces described by Zernike polynomials of different orders it was possible to simulate several target surfaces that may well resemble in vivo cases of refractive surgery candidates. An algorithm that uses the mathematical concept of convolution was implemented and several simulated surfaces were tested, ranging from low order

Observation of lens aberrations for high resolution electron microscopy II: Simple expressions for optimal estimates

Energy Technology Data Exchange (ETDEWEB)

Saxton, W. Owen, E-mail: wos1@cam.ac.uk

2015-04-15

This paper lists simple closed-form expressions estimating aberration coefficients (defocus, astigmatism, three-fold astigmatism, coma / misalignment, spherical aberration) on the basis of image shift or diffractogram shape measurements as a function of injected beam tilt. Simple estimators are given for a large number of injected tilt configurations, optimal in the sense of least-squares fitting of all the measurements, and so better than most reported previously. Standard errors are given for most, allowing different approaches to be compared. Special attention is given to the measurement of the spherical aberration, for which several simple procedures are given, and the effect of foreknowledge of this on other aberration estimates is noted. Details and optimal expressions are also given for a new and simple method of analysis, requiring measurements of the diffractogram mirror axis direction only, which are simpler to make than the focus and astigmatism measurements otherwise required. - Highlights: • Optimal estimators for CTEM lens aberrations are more accurate and/or use fewer observations. • Estimators have been found for defocus, astigmatism, three-fold astigmatism, coma and spherical aberration. • Estimators have been found relying on diffractogram shape, image shift and diffractogram orientation only, for a variety of beam tilts. • The standard error for each estimator has been found.

Microscopic validation of whole mouse micro-metastatic tumor imaging agents using cryo-imaging and sliding organ image registration

Science.gov (United States)

Liu, Yiqiao; Zhou, Bo; Qutaish, Mohammed; Wilson, David L.

2016-03-01

We created a metastasis imaging, analysis platform consisting of software and multi-spectral cryo-imaging system suitable for evaluating emerging imaging agents targeting micro-metastatic tumor. We analyzed CREKA-Gd in MRI, followed by cryo-imaging which repeatedly sectioned and tiled microscope images of the tissue block face, providing anatomical bright field and molecular fluorescence, enabling 3D microscopic imaging of the entire mouse with single metastatic cell sensitivity. To register MRI volumes to the cryo bright field reference, we used our standard mutual information, non-rigid registration which proceeded: preprocess --> affine --> B-spline non-rigid 3D registration. In this report, we created two modified approaches: mask where we registered locally over a smaller rectangular solid, and sliding organ. Briefly, in sliding organ, we segmented the organ, registered the organ and body volumes separately and combined results. Though sliding organ required manual annotation, it provided the best result as a standard to measure other registration methods. Regularization parameters for standard and mask methods were optimized in a grid search. Evaluations consisted of DICE, and visual scoring of a checkerboard display. Standard had accuracy of 2 voxels in all regions except near the kidney, where there were 5 voxels sliding. After mask and sliding organ correction, kidneys sliding were within 2 voxels, and Dice overlap increased 4%-10% in mask compared to standard. Mask generated comparable results with sliding organ and allowed a semi-automatic process.

Saturation and Energy Corrections for TeV Electrons and Photons

CERN Document Server

Clerbaux, Barbara; Mahmoud, Tariq; Marage, Pierre Edouard

2006-01-01

This note presents a study of the response of the CMS electromagnetic calorimeter ECAL to high energy electrons and photons (from 500 to 4000 GeV), using the full simulation of the CMS detector. The longitudinal containment and the lateral extension of high energy showers are discussed, and energy and eta dependent correction factors F(E_meas, eta), where E_meas = E_ECAL + E_HCAL, are determined in order to reconstruct the incident particle energy, using the energies measured in the ECAL and in the hadronic calorimeter HCAL. For ECAL barrel crystals with energy deposit higher than 1700 GeV, improvements are proposed to techniques aimed at correcting for the effects of electronics saturation.

Beam dynamics in rf guns and emittance correction techniques

International Nuclear Information System (INIS)

Serafini, L.

1994-01-01

In this paper we present a general review of beam dynamics in a laser-driven rf gun. The peculiarity of such an accelerating structure versus other conventional multi-cell linac structures is underlined on the basis of the Panofsky-Wenzel theorem, which is found to give a theoretical background for the well known Kim's model. A basic explanation for some proposed methods to correct rf induced emittance growth is also derived from the theorem. We also present three emittance correction techniques for the recovery of space-charge induced emittance growth, namely the optimum distributed disk-like bunch technique, the use of rf spatial harmonics to correct spherical aberration induced by space charge forces and the technique of emittance filtering by clipping the electron beam. The expected performances regarding the beam quality achievable with different techniques, as predicted by scaling laws and simulations, are analyzed, and, where available, compared to experimental results. (orig.)

Measurement of eye aberrations in a speckle field

International Nuclear Information System (INIS)

Larichev, A V; Ivanov, P V; Iroshnikov, N G; Shmalgauzen, V I

2001-01-01

The influence of speckles on the performance of a Shark-Hartmann wavefront sensor is investigated in the eye aberration studies. The dependence of the phase distortion measurement error on the characteristic speckle size is determined experimentally. Scanning of the reference source was used to suppress the speckle structure of the laser beam scattered by the retina. The technique developed by us made it possible to study the time dependence of the human eye aberrations with a resolution of 30 ms. (laser applications and other topics in quantum electronics)

Energetic particle induced desorption of water vapor cryo-condensate

International Nuclear Information System (INIS)

Menon, M.M.; Owen, L.W.; Simpkins, J.E.; Uckan, T.; Mioduszewski, P.K.

1990-01-01

An in-vessel cryo-condensation pump is being designed for the Advanced Divertor configuration of the DIII-D tokamak. To assess the importance of possible desorption of water vapor from the cryogenic surfaces of the pump due to impingement of energetic particles from the plasma, a 77 K surface on which a thin layer of water vapor was condensed was exposed to a tenuous plasma (density = 2 x 10 10 cm -3 , electron temperature = 3 eV). Significant desorption of the condensate occurred, suggesting that impingement of energeticparticles (10 eV) at flux levels of ∼10 16 cm 2 s -1 on cryogenic surfaces could potentially induce impurity problems in the tokamak plasma. A pumping configuration is presented in which this problem is minimized without sacrificing the pumping speed

D0 Cryo System Control System Autodialer

Energy Technology Data Exchange (ETDEWEB)

Urbin, J.; /Fermilab

1990-04-17

The DO cryogenic system is controlled by a TI565-PLC based control system. This allows the system to be unmanned when in steady state operation. System experts will need to be contacted when system parameters exceed normal operating points and reach alarm setpoints. The labwide FIRUS system provides one alarm monitor and communication link. An autodialer provides a second and more flexible alarm monitor and communication link. The autodialer monitors contact points in the control system and after receiving indication of an alarm accesses a list of experts which it calls until it receives an acknowledgement. There are several manufacturers and distributors of autodialer systems. This EN explains the search process the DO cryo group used to fmd an autodialer system that fit the cryo system's needs and includes information and specs for the unit we chose.

Assessment of refractive astigmatism and simulated therapeutic refractive surgery strategies in coma-like-aberrations-dominant corneal optics.

Science.gov (United States)

Zhou, Wen; Stojanovic, Aleksandar; Utheim, Tor Paaske

2016-01-01

The aim of the study is to raise the awareness of the influence of coma-like higher-order aberrations (HOAs) on power and orientation of refractive astigmatism (RA) and to explore how to account for that influence in the planning of topography-guided refractive surgery in eyes with coma-like-aberrations-dominant corneal optics. Eleven eyes with coma-like-aberrations-dominant corneal optics and with low lenticular astigmatism (LA) were selected for astigmatism analysis and for treatment simulations with topography-guided custom ablation. Vector analysis was used to evaluate the contribution of coma-like corneal HOAs to RA. Two different strategies were used for simulated treatments aiming to regularize irregular corneal optics: With both strategies correction of anterior corneal surface irregularities (corneal HOAs) were intended. Correction of total corneal astigmatism (TCA) and RA was intended as well with strategies 1 and 2, respectively. Axis of discrepant astigmatism (RA minus TCA minus LA) correlated strongly with axis of coma. Vertical coma influenced RA by canceling the effect of the with-the-rule astigmatism and increasing the effect of the against-the-rule astigmatism. After simulated correction of anterior corneal HOAs along with TCA and RA (strategies 1 and 2), only a small amount of anterior corneal astigmatism (ACA) and no TCA remained after strategy 1, while considerable amount of ACA and TCA remained after strategy 2. Coma-like corneal aberrations seem to contribute a considerable astigmatic component to RA in eyes with coma-like-aberrations dominant corneal optics. If topography-guided ablation is programmed to correct the corneal HOAs and RA, the astigmatic component caused by the coma-like corneal HOAs will be treated twice and will result in induced astigmatism. Disregarding RA and treating TCA along with the corneal HOAs is recommended instead.

Chromosomal aberrations and DNA damage in human populations exposed to the processing of electronics waste.

Science.gov (United States)

Liu, Qiang; Cao, Jia; Li, Ke Qiu; Miao, Xu Hong; Li, Guang; Fan, Fei Yue; Zhao, Yong Cheng

2009-05-01

It has been known that the pollutants of electronic wastes (E-wastes) can lead to severe pollution to the environment. It has been reported that about 50% to 80% of E-wastes from developed countries are exported to Asia and Africa. It has become a major global environmental problem to deal with 'E-wastes'. E-waste recycling has remained primitive in Jinghai, China. This not only produces enormous environmental pollution but also can bring about toxic or genotoxic effects on the human body, threatening the health of both current residents and future generations living in the local environment. The concentration of lead in the blood of children in the E-waste polluted area in China is higher than that of the control area. But little is known about the cytogenetic effect to human beings caused by the pollution of E-wastes. In the present study, experiments have been performed to investigate the genetics of permanent residents of three villages with numerous E-waste disposal sites and to analyze the harmful effects of exposure to E-wastes. In total, 171 villagers (exposed group) were randomly selected from permanent residents of three villages located in Jinghai County of Tianjin, China, where there has been massive disposal of E-wastes. Thirty villagers were selected from the neighboring towns without E-waste disposal sites to serve as controls. Chromosomal aberrations and cytokinesis blocking micronucleus were performed to detect the cytogenetic effect, dic + r (dicentric and ring chromosome), monomer, fragments (acentric fragments, minute chromosomes, and acentric rings), translocation, satellite, quadriradial, total aberrations, and micronuclear rate were scored for each subject. DNA damage was detected using comet assay; the DNA percentage in the comet tail (TDNA%), tail moment (TM), and Olive tail moment (OTM) were recorded to describe DNA damage to lymphocytes. The total chromosome aberration rates (5.50%) and micronuclear rates (16.99%) of the exposure group

Transmission Electron Microscopy Physics of Image Formation

CERN Document Server

Kohl, Helmut

2008-01-01

Transmission Electron Microscopy: Physics of Image Formation presents the theory of image and contrast formation, and the analytical modes in transmission electron microscopy. The principles of particle and wave optics of electrons are described. Electron-specimen interactions are discussed for evaluating the theory of scattering and phase contrast. Also discussed are the kinematical and dynamical theories of electron diffraction and their applications for crystal-structure analysis and imaging of lattices and their defects. X-ray microanalysis and electron energy-loss spectroscopy are treated as analytical methods. Specimen damage and contamination by electron irradiation limits the resolution for biological and some inorganic specimens. This fifth edition includes discussion of recent progress, especially in the area of aberration correction and energy filtering; moreover, the topics introduced in the fourth edition have been updated. Transmission Electron Microscopy: Physics of Image Formation is written f...

Measuring chromatic aberrations in imaging systems using plasmonic nanoparticles.

Science.gov (United States)

Gennaro, Sylvain D; Roschuk, Tyler R; Maier, Stefan A; Oulton, Rupert F

2016-04-01

We demonstrate a method to measure chromatic aberrations of microscope objectives with metallic nanoparticles using white light. Extinction spectra are recorded while scanning a single nanoparticle through a lens's focal plane. We show a direct correlation between the focal wavelength and the longitudinal chromatic focal shift through our analysis of the variations between the scanned extinction spectra at each scan position and the peak extinction over the entire scan. The method has been tested on achromat and apochromat objectives using aluminum disks varying in size from 260-520 nm. Our method is straightforward, robust, low cost, and broadband with a sensitivity suitable for assessing longitudinal chromatic aberrations in high-numerical-aperture apochromatic corrected lenses.

CryoSat-2 Processing and Model Interpretation of Greenland Ice Sheet Volume Changes

Science.gov (United States)

Nilsson, J.; Gardner, A. S.; Sandberg Sorensen, L.

2015-12-01

CryoSat-2 was launched in late 2010 tasked with monitoring the changes of the Earth's land and sea ice. It carries a novel radar altimeter allowing the satellite to monitor changes in highly complex terrain, such as smaller ice caps, glaciers and the marginal areas of the ice sheets. Here we present on the development and validation of an independent elevation retrieval processing chain and respective elevation changes based on ESA's L1B data. Overall we find large improvement in both accuracy and precision over Greenland relative to ESA's L2 product when comparing against both airborne data and crossover analysis. The seasonal component and spatial sampling of the surface elevation changes where also compared against ICESat derived changes from 2003-2009. The comparison showed good agreement between the to product on a local scale. However, a global sampling bias was detected in the seasonal signal due to the clustering of CryoSat-2 data in higher elevation areas. The retrieval processing chain presented here does not correct for changes in surface scattering conditions and appears to be insensitive to the 2012 melt event (Nilsson et al., 2015). This in contrast to the elevation changes derived from ESA's L2 elevation product, which where found to be sensitive to the effects of the melt event. The positive elevation bias created by the event introduced a discrepancy between the two products with a magnitude of roughly 90 km3/year. This difference can directly be attributed to the differences in retracking procedure pointing to the importance of the retracking of the radar waveforms for altimetric volume change studies. Greenland 2012 melt event effects on CryoSat-2 radar altimetry./ Nilsson, Johan; Vallelonga, Paul Travis; Simonsen, Sebastian Bjerregaard; Sørensen, Louise Sandberg; Forsberg, René; Dahl-Jensen, Dorthe; Hirabayashi, Motohiro; Goto-Azuma, Kumiko; Hvidberg, Christine S.; Kjær, Helle A.; Satow, Kazuhide.

Environmental TEM in an Aberration Corrected Microscope

DEFF Research Database (Denmark)

Hansen, Thomas Willum; Wagner, Jakob Birkedal

‐resolution imaging. A gaseous atmosphere in the pole‐piece gap of the objective lens of the microscope alters both the incoming electron wave prior to interaction with the sample and the outgoing wave below the sample. Whereas conventional TEM samples are usually thin (below 10‐20 nm), the gas in the environmental...... the microscope column. The effects of gas on the electron wave in the objective lens are not well understood and needs further attention. Imaging samples with a simple geometry, such as gold particles on a flat graphene substrate and analyzing the variations in contrast, provides a means for understanding...... results from imaging in various elemental as well as di‐molecular gases and their effect on imaging and spectroscopy in the environmental transmission electron microscope....

Cryo-EM structure of the E. coli translating ribosome in complex with SRP and its receptor

Science.gov (United States)

Estrozi, Leandro F.; Boehringer, Daniel; Shan, Shu-ou; Ban, Nenad; Schaffitzel, Christiane

2013-01-01

We report the early conformation of the E. coli signal recognition particle (SRP) and its receptor FtsY bound to the translating ribosome by cryo-electron microscopy. FtsY binds to the tetraloop of the SRP RNA whereas the NG-domains of the SRP protein and FtsY interact weakly in this conformation. Our results suggest that optimal positioning of the SRP RNA tetraloop and the Ffh NG-domain leads to FtsY recruitment. PMID:21151118

Electrostatic axisymmetric mirror with removable spherical aberration

International Nuclear Information System (INIS)

Birmuzaev, S.B.; Serikbaeva, G.S.; Hizirova, M.A.

1999-01-01

The electrostatic axisymmetric mirror, assembled from three coaxial cylinders with an equal diameter d and under the potential v1, v2 and v3, was computed. The proportions of geometrical and electric parameters of the mirror, with which the spherical 3-order aberration may be eliminated, were determined. The computation outcomes of the case, when the focal power of the mirror is enough large and the object plane in the focus is out of its field, are presented (Fig. 1 - potentials proportion that makes elimination of the spherical aberration possible; Fig. 2 - the focus coordinates when the spherical aberration is eliminated). The geometrical values are presented by d, and the electric ones are presented by v1. The figures on the curves present a length of the second (middle) electrode. The zero point is located in the middle of the gap between the first and second electrodes The investigated mirror may be used as a lens for the transmission electron microscope

Space Active Optics: toward optimized correcting mirrors for future large spaceborne observatories

Science.gov (United States)

Laslandes, Marie; Hugot, Emmanuel; Ferrari, Marc; Lemaitre, Gérard; Liotard, Arnaud

2011-10-01

Wave-front correction in optical instruments is often needed, either to compensate Optical Path Differences, off-axis aberrations or mirrors deformations. Active optics techniques are developed to allow efficient corrections with deformable mirrors. In this paper, we will present the conception of particular deformation systems which could be used in space telescopes and instruments in order to improve their performances while allowing relaxing specifications on the global system stability. A first section will be dedicated to the design and performance analysis of an active mirror specifically designed to compensate for aberrations that might appear in future 3m-class space telescopes, due to lightweight primary mirrors, thermal variations or weightless conditions. A second section will be dedicated to a brand new design of active mirror, able to compensate for given combinations of aberrations with a single actuator. If the aberrations to be corrected in an instrument and their evolutions are known in advance, an optimal system geometry can be determined thanks to the elasticity theory and Finite Element Analysis.

Automatic phase aberration compensation for digital holographic microscopy based on deep learning background detection.

Science.gov (United States)

Nguyen, Thanh; Bui, Vy; Lam, Van; Raub, Christopher B; Chang, Lin-Ching; Nehmetallah, George

2017-06-26

We propose a fully automatic technique to obtain aberration free quantitative phase imaging in digital holographic microscopy (DHM) based on deep learning. The traditional DHM solves the phase aberration compensation problem by manually detecting the background for quantitative measurement. This would be a drawback in real time implementation and for dynamic processes such as cell migration phenomena. A recent automatic aberration compensation approach using principle component analysis (PCA) in DHM avoids human intervention regardless of the cells' motion. However, it corrects spherical/elliptical aberration only and disregards the higher order aberrations. Traditional image segmentation techniques can be employed to spatially detect cell locations. Ideally, automatic image segmentation techniques make real time measurement possible. However, existing automatic unsupervised segmentation techniques have poor performance when applied to DHM phase images because of aberrations and speckle noise. In this paper, we propose a novel method that combines a supervised deep learning technique with convolutional neural network (CNN) and Zernike polynomial fitting (ZPF). The deep learning CNN is implemented to perform automatic background region detection that allows for ZPF to compute the self-conjugated phase to compensate for most aberrations.

Biological applications of phase-contrast electron microscopy.

Science.gov (United States)

Nagayama, Kuniaki

2014-01-01

Here, I review the principles and applications of phase-contrast electron microscopy using phase plates. First, I develop the principle of phase contrast based on a minimal model of microscopy, introducing a double Fourier-transform process to mathematically formulate the image formation. Next, I explain four phase-contrast (PC) schemes, defocus PC, Zernike PC, Hilbert differential contrast, and schlieren optics, as image-filtering processes in the context of the minimal model, with particular emphases on the Zernike PC and corresponding Zernike phase plates. Finally, I review applications of Zernike PC cryo-electron microscopy to biological systems such as protein molecules, virus particles, and cells, including single-particle analysis to delineate three-dimensional (3D) structures of protein and virus particles and cryo-electron tomography to reconstruct 3D images of complex protein systems and cells.

The coastal mean dynamic topography in Norway observed by CryoSat-2 and GOCE

DEFF Research Database (Denmark)

Idžanović, Martina; Ophaug, Vegard; Andersen, Ole Baltazar

2017-01-01

New-generation synthetic aperture radar altimetry, as implemented on CryoSat-2, observes sea surface heights in coastal areas that were previously not monitored by conventional altimetry. Therefore,CryoSat-2 is expected to improve the coastal mean dynamic topography (MDT). However, the MDT remains...

The crystal structure of a coxsackievirus B3-RD variant and a refined 9-angstrom cryo-electron microscopy reconstruction of the virus complexed with decay-accelerating factor (DAF) provide a new footprint of DAF on the virus surface.

Science.gov (United States)

Yoder, Joshua D; Cifuente, Javier O; Pan, Jieyan; Bergelson, Jeffrey M; Hafenstein, Susan

2012-12-01

The coxsackievirus-adenovirus receptor (CAR) and decay-accelerating factor (DAF) have been identified as cellular receptors for coxsackievirus B3 (CVB3). The first described DAF-binding isolate was obtained during passage of the prototype strain, Nancy, on rhabdomyosarcoma (RD) cells, which express DAF but very little CAR. Here, the structure of the resulting variant, CVB3-RD, has been solved by X-ray crystallography to 2.74 Å, and a cryo-electron microscopy reconstruction of CVB3-RD complexed with DAF has been refined to 9.0 Å. This new high-resolution structure permits us to correct an error in our previous view of DAF-virus interactions, providing a new footprint of DAF that bridges two adjacent protomers. The contact sites between the virus and DAF clearly encompass CVB3-RD residues recently shown to be required for binding to DAF; these residues interact with DAF short consensus repeat 2 (SCR2), which is known to be essential for virus binding. Based on the new structure, the mode of the DAF interaction with CVB3 differs significantly from the mode reported previously for DAF binding to echoviruses.

Reduced sintering of mass-selected Au clusters on SiO₂ by alloying with Ti: an aberration-corrected STEM and computational study

DEFF Research Database (Denmark)

Niu, Yubiao; Schlexer, Philomena; Sebök, Béla

2018-01-01

with a reactive metal, Ti. Mass-selected Au/Ti clusters (400 000 amu) and Au2057 clusters (405 229 amu) were produced with a magnetron sputtering, gas condensation cluster beam source in conjunction with a lateral time-of-flight mass filter, deposited onto a silica support and characterised by XPS and LEIS....... The sintering dynamics of mass-selected Au and Au/Ti alloy nanoclusters were investigated in real space and real time with atomic resolution aberration-corrected HAADF-STEM imaging, supported by model DFT calculations. A strong anchoring effect was revealed in the case of the Au/Ti clusters, because of a much...

Electron dose dependence of signal-to-noise ratio, atom contrast and resolution in transmission electron microscope images

International Nuclear Information System (INIS)

Lee, Z.; Rose, H.; Lehtinen, O.; Biskupek, J.; Kaiser, U.

2014-01-01

In order to achieve the highest resolution in aberration-corrected (AC) high-resolution transmission electron microscopy (HRTEM) images, high electron doses are required which only a few samples can withstand. In this paper we perform dose-dependent AC-HRTEM image calculations, and study the dependence of the signal-to-noise ratio, atom contrast and resolution on electron dose and sampling. We introduce dose-dependent contrast, which can be used to evaluate the visibility of objects under different dose conditions. Based on our calculations, we determine optimum samplings for high and low electron dose imaging conditions. - Highlights: • The definition of dose-dependent atom contrast is introduced. • The dependence of the signal-to-noise ratio, atom contrast and specimen resolution on electron dose and sampling is explored. • The optimum sampling can be determined according to different dose conditions

CryoSat SIRAL: Instrument Performance After 5 Years of Operations

Science.gov (United States)

Scagliola, Michele; Fornari, Marco; Bouffard, Jerome; Parrinello, Tommaso

2016-08-01

CryoSat's Synthetic Interferometric Radar Altimeter (SIRAL) [1] is a Ku-band pulsewidth limited radar altimeter that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for Delay/Doppler processing [2]. Moreover SIRAL takes advantage of two antennas mounted across-track for interferometric capability, in order to determine the across-track direction from which the echo is received [3].The calibration strategy for SIRAL includes both internal calibrations and external calibration [1,7]. Due to the fact that SIRAL is an interferometric phase coherent pulse-width limited radar altimeter, a proper calibration approach has been developed. In this paper we will describe as first the internal calibration strategy and then the different calibration corrections that are applied to science data. The internal calibration results over more than five years of mission will be presented, analysing their temporal evolution in order to highlight the stability of the instrument over its life.

Cryo-EM structures of the mammalian endo-lysosomal TRPML1 channel elucidate the combined regulation mechanism

Directory of Open Access Journals (Sweden)

Sensen Zhang

2017-09-01

Full Text Available Abstract TRPML1 channel is a non-selective group-2 transient receptor potential (TRP channel with Ca2+ permeability. Located mainly in late endosome and lysosome of all mammalian cell types, TRPML1 is indispensable in the processes of endocytosis, membrane trafficking, and lysosome biogenesis. Mutations of TRPML1 cause a severe lysosomal storage disorder called mucolipidosis type IV (MLIV. In the present study, we determined the cryo-electron microscopy (cryo-EM structures of Mus musculus TRPML1 (mTRPML1 in lipid nanodiscs and Amphipols. Two distinct states of mTRPML1 in Amphipols are added to the closed state, on which could represent two different confirmations upon activation and regulation. The polycystin-mucolipin domain (PMD may sense the luminal/extracellular stimuli and undergo a “move upward” motion during endocytosis, thus triggering the overall conformational change in TRPML1. Based on the structural comparisons, we propose TRPML1 is regulated by pH, Ca2+, and phosphoinositides in a combined manner so as to accommodate the dynamic endocytosis process.

Deep learning-based subdivision approach for large scale macromolecules structure recovery from electron cryo tomograms.

Science.gov (United States)

Xu, Min; Chai, Xiaoqi; Muthakana, Hariank; Liang, Xiaodan; Yang, Ge; Zeev-Ben-Mordehai, Tzviya; Xing, Eric P

2017-07-15

Cellular Electron CryoTomography (CECT) enables 3D visualization of cellular organization at near-native state and in sub-molecular resolution, making it a powerful tool for analyzing structures of macromolecular complexes and their spatial organizations inside single cells. However, high degree of structural complexity together with practical imaging limitations makes the systematic de novo discovery of structures within cells challenging. It would likely require averaging and classifying millions of subtomograms potentially containing hundreds of highly heterogeneous structural classes. Although it is no longer difficult to acquire CECT data containing such amount of subtomograms due to advances in data acquisition automation, existing computational approaches have very limited scalability or discrimination ability, making them incapable of processing such amount of data. To complement existing approaches, in this article we propose a new approach for subdividing subtomograms into smaller but relatively homogeneous subsets. The structures in these subsets can then be separately recovered using existing computation intensive methods. Our approach is based on supervised structural feature extraction using deep learning, in combination with unsupervised clustering and reference-free classification. Our experiments show that, compared with existing unsupervised rotation invariant feature and pose-normalization based approaches, our new approach achieves significant improvements in both discrimination ability and scalability. More importantly, our new approach is able to discover new structural classes and recover structures that do not exist in training data. Source code freely available at http://www.cs.cmu.edu/∼mxu1/software . mxu1@cs.cmu.edu. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Correct Brillouin zone and electronic structure of BiPd

Science.gov (United States)

Yaresko, Alexander; Schnyder, Andreas P.; Benia, Hadj M.; Yim, Chi-Ming; Levy, Giorgio; Damascelli, Andrea; Ast, Christian R.; Peets, Darren C.; Wahl, Peter

2018-02-01

A promising route to the realization of Majorana fermions is in noncentrosymmetric superconductors, in which spin-orbit coupling lifts the spin degeneracy of both bulk and surface bands. A detailed assessment of the electronic structure is critical to evaluate their suitability for this through establishing the topological properties of the electronic structure. This requires correct identification of the time-reversal-invariant momenta. One such material is BiPd, a recently rediscovered noncentrosymmetric superconductor which can be grown in large, high-quality single crystals and has been studied by several groups using angular resolved photoemission to establish its surface electronic structure. Many of the published electronic structure studies on this material are based on a reciprocal unit cell which is not the actual Brillouin zone of the material. We show here the consequences of this for the electronic structures and show how the inferred topological nature of the material is affected.

A Novel Low Energy Electron Microscope for DNA Sequencing and Surface Analysis

Science.gov (United States)

Mankos, M.; Shadman, K.; Persson, H.H.J.; N’Diaye, A.T.; Schmid, A.K.; Davis, R.W.

2014-01-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel technique that is directed towards imaging nanostructures and surfaces with sub-nanometer resolution. The technique combines a monochromator, a mirror aberration corrector, an energy filter, and dual beam illumination in a single instrument. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. Simulation results predict that the novel aberration corrector design will eliminate the second rank chromatic and third and fifth order spherical aberrations, thereby improving the resolution into the sub-nanometer regime at landing energies as low as one hundred electron-Volts. The energy filter produces a beam that can extract detailed information about the chemical composition and local electronic states of non-periodic objects such as nanoparticles, interfaces, defects, and macromolecules. The dual flood illumination eliminates charging effects that are generated when a conventional LEEM is used to image insulating specimens. A potential application for MAD-LEEM is in DNA sequencing, which requires high resolution to distinguish the individual bases and high speed to reduce the cost. The MAD-LEEM approach images the DNA with low electron impact energies, which provides nucleobase contrast mechanisms without organometallic labels. Furthermore, the micron-size field of view when combined with imaging on the fly provides long read lengths, thereby reducing the demand on assembling the sequence. Experimental results from bulk specimens with immobilized single-base oligonucleotides demonstrate that base specific contrast is available with reflected, photo-emitted, and Auger electrons. Image contrast simulations of model rectangular features mimicking the individual nucleotides in a DNA strand have been developed to translate measurements of contrast on bulk DNA to the detectability of

A novel low energy electron microscope for DNA sequencing and surface analysis.

Science.gov (United States)

Mankos, M; Shadman, K; Persson, H H J; N'Diaye, A T; Schmid, A K; Davis, R W

2014-10-01

Monochromatic, aberration-corrected, dual-beam low energy electron microscopy (MAD-LEEM) is a novel technique that is directed towards imaging nanostructures and surfaces with sub-nanometer resolution. The technique combines a monochromator, a mirror aberration corrector, an energy filter, and dual beam illumination in a single instrument. The monochromator reduces the energy spread of the illuminating electron beam, which significantly improves spectroscopic and spatial resolution. Simulation results predict that the novel aberration corrector design will eliminate the second rank chromatic and third and fifth order spherical aberrations, thereby improving the resolution into the sub-nanometer regime at landing energies as low as one hundred electron-Volts. The energy filter produces a beam that can extract detailed information about the chemical composition and local electronic states of non-periodic objects such as nanoparticles, interfaces, defects, and macromolecules. The dual flood illumination eliminates charging effects that are generated when a conventional LEEM is used to image insulating specimens. A potential application for MAD-LEEM is in DNA sequencing, which requires high resolution to distinguish the individual bases and high speed to reduce the cost. The MAD-LEEM approach images the DNA with low electron impact energies, which provides nucleobase contrast mechanisms without organometallic labels. Furthermore, the micron-size field of view when combined with imaging on the fly provides long read lengths, thereby reducing the demand on assembling the sequence. Experimental results from bulk specimens with immobilized single-base oligonucleotides demonstrate that base specific contrast is available with reflected, photo-emitted, and Auger electrons. Image contrast simulations of model rectangular features mimicking the individual nucleotides in a DNA strand have been developed to translate measurements of contrast on bulk DNA to the detectability of

Population spherical aberration: associations with ametropia, age, corneal curvature, and image quality

Directory of Open Access Journals (Sweden)

Kingston AC

2013-05-01

Full Text Available Amanda C Kingston,1,2 Ian G Cox11Bausch + Lomb, Rochester, NY, USA; 2Department of Biomedical Engineering, University of Rochester, Rochester, NY, USAPurpose: The aim of this analysis was to determine the total ocular wavefront aberration values of a large phakic population of physiologically normal, ametropic eyes, gathered under the same clinical protocol using the same diagnostic wavefront sensor.Materials and methods: Studies were conducted at multiple sites in Asia, North America, Europe, and Australia. A Bausch + Lomb Zywave II Wavefront Aberrometer (Rochester, NY, USA was used to measure the lower and higher order aberrations of each eye. Data analysis was conducted using linear regression analysis to determine the relationship between total spherical aberration, ametropia, age, corneal curvature, and image quality.Results: Linear regression analysis showed no correlation (r = 0.0207, P = 0.4874 between degree of ametropia and the amount of spherical aberration. There was also no correlation when the population was stratified into myopic and hyperopic refractive groups (rm = 0.0529, Pm = 0.0804 and rh = 0.1572, Ph = 0.2754. There was a statistically significant and weak positive correlation (r = 0.1962, P < 0.001 between age and the amount of spherical aberration measured in the eye; spherical aberration became more positive with increasing age. Also, there was a statistically significant and moderately positive correlation (r = 0.3611, P < 0.001 with steepness of corneal curvature; spherical aberration became more positive with increasing power of the anterior corneal surface. Assessment of image quality using optical design software (Zemax™, Bellevue, WA, USA showed that there was an overall benefit in correcting the average spherical aberration of this population.Conclusion: Analysis of this dataset provides insights into the inherent spherical aberration of a typical phakic, pre-presbyopic, population and provides the ability to

Digital holography with electron wave: measuring into the nanoworld

Science.gov (United States)

Mendoza Santoyo, Fernando; Voelkl, Edgar

2016-04-01

Dennis Gabor invented Holography in 1949. His main concern at the time was centered on the spherical aberration correction in the recently created electron microscopes, especially after O. Scherzer had shown mathematically that round electron optical lenses always have a positive spherical aberration coefficient and the mechanical requirements for minimizing the spherical aberration were too high to allow for atomic resolution. At the time the lack of coherent electron sources meant that in-line holography was developed using quasi-coherent light sources. As such Holography did not produce scientific good enough results to be considered a must use tool. In 1956, G. Moellenstedt invented a device called a wire-biprism that allowed the object and reference beams to be combined in an off-axis configuration. The invention of the laser at the end of the 1950s gave a great leap to Holography since this light source was highly coherent and hence led to the invention of Holographic Interferometry during the first lustrum of the 1960s. This new discipline in the Optics field has successfully evolved to become a trusted tool in a wide variety of areas. Coherent electron sources were made available only by the late 1970s, a fact that gave an outstanding impulse to electron holography so that today nanomaterials and structures belonging to a wide variety of subjects can be characterized in regards to their physical and mechanical parameters. This invited paper will present and discuss electron holography's state of the art applications to study the shape of nanoparticles and bacteria, and the qualitative and quantitative study of magnetic and electric fields produced by novel nano-structures.

Aberrant chlamydial developmental forms in the gastrointestinal tract of pigs spontaneously and experimentally infected with Chlamydia suis.

Science.gov (United States)

Pospischil, Andreas; Borel, Nicole; Chowdhury, Emdad H; Guscetti, Franco

2009-03-16

The phenomenon of persistence is well known from in vitro studies, where it is associated with the production of aberrant bodies, but its occurrence in vivo is less well documented. The objective of this study was to search for aberrant bodies in intestinal tissues from pigs, describe their ultrastructure, and investigate the suitability of immunohistochemical staining for chlamydial heat shock protein 60 (cHSP60) to detect such forms. Intestinal tissues derived from pigs naturally and experimentally infected with Chlamydia (C.) suis were examined by immunohistochemistry, transmission electron microscopy and immunogold electron microscopy. The chlamydial species involved in the natural infection were determined using an Array Tube Microarray to C. suis and Chlamydophila abortus. Ultrastructurally, aberrant bodies were detected in the gut of both naturally and experimentally infected pigs. Immunogold electron microscopy showed that the aberrant bodies were labeled less strongly than the normal forms by antibodies against LPS and cHSP60 respectively. It was concluded that aberrant bodies occur in vivo in pigs and that the gnotobiotic pig model might be suitable for the study of chlamydial persistence in vivo. The antibody against cHSP60 does not appear to be suitable to specifically detect such forms.

CryoSat-2 radar altimetry for monitoring freshwater resources of China

DEFF Research Database (Denmark)

Jiang, Liguang; Nielsen, Karina; Andersen, Ole Baltazar

2017-01-01

, the Northeast China Plain, and the central Yangtze River basin. Estimated changes in volume indicate that surface water variation contributes significantly to terrestrial storage variation, especially in the Qaidam Basin and the Tibetan Plateau. CryoSat-2 is capable of measuring regional-scale river level....... Comparatively, accuracy is much lower over the Yangtze and Pearl Rivers (RMSE ~ 2.6 m and ~ 3.3 m), probably due to intensive inland waterway navigation. CryoSat-2 shows great potential for monitoring surface water at national scale in China....

Effect of Spherical Aberration on the Optical Quality after Implantation of Two Different Aspherical Intraocular Lenses

Directory of Open Access Journals (Sweden)

Michael Lasta

2017-01-01

Full Text Available Purpose. To compare the effect of spherical aberration on optical quality in eyes with two different aspherical intraocular lenses. Methods. 120 eyes of 60 patients underwent phacoemulsification. In patients’ eyes, an aberration-free IOL (Aspira-aA; Human Optics or an aberration-correcting aspherical IOL (Tecnis ZCB00; Abott Medical Optics was randomly implanted. After surgery, contrast sensitivity and wavefront measurements as well as tilt and decentration measurements were performed. Results. Contrast sensitivity was significantly higher in eyes with Aspira lens under mesopic conditions with 12 cycles per degree (CPD and under photopic conditions with 18 CPD (p=0.02. Wavefront measurements showed a higher total spherical aberration with a minimal pupil size of 4 mm in the Aspira group (0.05 ± 0.03 than in the Tecnis group (0.03 ± 0.02 (p=0.001. Strehl ratio was higher in eyes with Tecnis (0.28 ± 0.17 with a minimal pupil size larger than 5 mm than that with Aspira (0.16 ± 0.14 (p=0.04. In pupils with a minimum diameter of 4 mm spherical aberration had a significant effect on Strehl ratio, but not in pupils with a diameter less than 4 mm. Conclusions. Optical quality was better in eyes with the aberration-correcting Tecnis IOL when pupils were large. In contrast, this could not be shown in eyes with pupils under 4 mm or larger. This trial is registered with Clinicaltrials.gov NCT03224728.

Cryo-EM structures of the TMEM16A calcium-activated chloride channel.

Science.gov (United States)

Dang, Shangyu; Feng, Shengjie; Tien, Jason; Peters, Christian J; Bulkley, David; Lolicato, Marco; Zhao, Jianhua; Zuberbühler, Kathrin; Ye, Wenlei; Qi, Lijun; Chen, Tingxu; Craik, Charles S; Jan, Yuh Nung; Minor, Daniel L; Cheng, Yifan; Jan, Lily Yeh

2017-12-21

Calcium-activated chloride channels (CaCCs) encoded by TMEM16A control neuronal signalling, smooth muscle contraction, airway and exocrine gland secretion, and rhythmic movements of the gastrointestinal system. To understand how CaCCs mediate and control anion permeation to fulfil these physiological functions, knowledge of the mammalian TMEM16A structure and identification of its pore-lining residues are essential. TMEM16A forms a dimer with two pores. Previous CaCC structural analyses have relied on homology modelling of a homologue (nhTMEM16) from the fungus Nectria haematococca that functions primarily as a lipid scramblase, as well as subnanometre-resolution electron cryo-microscopy. Here we present de novo atomic structures of the transmembrane domains of mouse TMEM16A in nanodiscs and in lauryl maltose neopentyl glycol as determined by single-particle electron cryo-microscopy. These structures reveal the ion permeation pore and represent different functional states. The structure in lauryl maltose neopentyl glycol has one Ca 2+ ion resolved within each monomer with a constricted pore; this is likely to correspond to a closed state, because a CaCC with a single Ca 2+ occupancy requires membrane depolarization in order to open (C.J.P. et al., manuscript submitted). The structure in nanodiscs has two Ca 2+ ions per monomer and its pore is in a closed conformation; this probably reflects channel rundown, which is the gradual loss of channel activity that follows prolonged CaCC activation in 1 mM Ca 2+ . Our mutagenesis and electrophysiological studies, prompted by analyses of the structures, identified ten residues distributed along the pore that interact with permeant anions and affect anion selectivity, as well as seven pore-lining residues that cluster near pore constrictions and regulate channel gating. Together, these results clarify the basis of CaCC anion conduction.

Fast and sensitive rigid-body fitting into cryo-EM density maps with PowerFit

NARCIS (Netherlands)

C.p.van Zundert, Gydo; M.j.j. Bonvin, Alexandre

2015-01-01

Cryo-EM is a rapidly developing method to investigate the three dimensional structure of large macromolecular complexes. In spite of all the advances in the field, the resolution of most cryo-EM density maps is too low for de novo model building. Therefore, the data are often complemented by fitting

Comparing elevation and freeboard from IceBridge and four different CryoSat-2 retrackers for coincident sea ice observations

Science.gov (United States)

Yi, D.; Kurtz, N. T.; Harbeck, J.

2017-12-01

The airborne IceBridge and spaceborne Cryosat-2 missions observe polar sea ice at different altitudes with different footprint sizes and often at different time and locations. Many studies use different retrackers to derive Cryosat-2 surface elevation, which we find causes large differences in the elevation and freeboard comparisons of IceBridge and Cryosat-2. In this study, we compare sea ice surface elevation and freeboard using 8 coincident CryoSat-2, ATM, and LVIS observations with IceBridge airplanes under flying the Cryosat-2 ground tracks. We apply identical ellipsoid, geoid model, tide model, and atmospheric correction to CryoSat-2 and IceBridge data to reduce elevation bias due to their differences. IceBridge's ATM and LVIS elevation and freeboard and Snow Radar snow depth are averaged at each CryoSat-2 footprint for comparison. The four different Cryosat-2 retrackers (ESA, GSFC, AWI, and JPL) show distinct differences in mean elevation up to 0.35 meters over leads and over floes, which suggests that systematic elevation bias exists between the retrackers. The mean IceBridge elevation over leads is within the mean elevation distribution of the four Cryosat-2 retrackers. The mean IceBridge elevation over floes is above the mean elevation distribution of the four Cryosat-2 retrackers. After removing the snow depth from IceBridge elevation, over floe, the mean elevation of IceBridge is within the mean elevation distribution of the four Cryosat-2 retrackers. By identifying the strengths and weaknesses of the retrackers, this study provides a mechanism to improve freeboard retrievals from existing methods.

OMNY—A tOMography Nano crYo stage

Science.gov (United States)

Holler, M.; Raabe, J.; Diaz, A.; Guizar-Sicairos, M.; Wepf, R.; Odstrcil, M.; Shaik, F. R.; Panneels, V.; Menzel, A.; Sarafimov, B.; Maag, S.; Wang, X.; Thominet, V.; Walther, H.; Lachat, T.; Vitins, M.; Bunk, O.

2018-04-01

For many scientific questions gaining three-dimensional insight into a specimen can provide valuable information. We here present an instrument called "tOMography Nano crYo (OMNY)," dedicated to high resolution 3D scanning x-ray microscopy at cryogenic conditions via hard X-ray ptychography. Ptychography is a lens-less imaging method requiring accurate sample positioning. In OMNY, this in achieved via dedicated laser interferometry and closed-loop position control reaching sub-10 nm positioning accuracy. Cryogenic sample conditions are maintained via conductive cooling. 90 K can be reached when using liquid nitrogen as coolant, and 10 K is possible with liquid helium. A cryogenic sample-change mechanism permits measurements of cryogenically fixed specimens. We compare images obtained with OMNY with older measurements performed using a nitrogen gas cryo-jet of stained, epoxy-embedded retina tissue and of frozen-hydrated Chlamydomonas cells.

Higher order aberrations in amblyopic children and their role in refractory amblyopia

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Arnaldo Dias-Santos

2014-12-01

Full Text Available Objective: Some studies have hypothesized that an unfavourable higher order aberrometric profile could act as an amblyogenic mechanism and may be responsible for some amblyopic cases that are refractory to conventional treatment or cases of “idiopathic” amblyopia. This study compared the aberrometric profile in amblyopic children to that of children with normal visual development and compared the aberrometric profile in corrected amblyopic eyes and refractory amblyopic eyes with that of healthy eyes. Methods: Cross-sectional study with three groups of children – the CA group (22 eyes of 11 children with unilateral corrected amblyopia, the RA group (24 eyes of 13 children with unilateral refractory amblyopia and the C group (28 eyes of 14 children with normal visual development. Higher order aberrations were evaluated using an OPD-Scan III (NIDEK. Comparisons of the aberrometric profile were made between these groups as well as between the amblyopic and healthy eyes within the CA and RA groups. Results: Higher order aberrations with greater impact in visual quality were not significantly higher in the CA and RA groups when compared with the C group. Moreover, there were no statistically significant differences in the higher order aberrometric profile between the amblyopic and healthy eyes within the CA and RA groups. Conclusions: Contrary to lower order aberrations (e.g., myopia, hyperopia, primary astigmatism, higher order aberrations do not seem to be involved in the etiopathogenesis of amblyopia. Therefore, these are likely not the cause of most cases of refractory amblyopia.

Cryo-STEM-EDX spectroscopy for the characterisation of nanoparticles in cell culture media

Science.gov (United States)

Ilett, M.; Bamiduro, F.; Matar, O.; Brown, A.; Brydson, R.; Hondow, N.

2017-09-01

We present a study of barium titanate nanoparticles dispersed in cell culture media. Scanning transmission electron microscopy combined with energy dispersive X-ray spectroscopy was undertaken on samples prepared using both conventional drop casting and also plunge freezing and examination under cryogenic conditions. This showed that drying artefacts occurred during conventional sample preparation, whereby some salt components of the cell culture media accumulated around the barium titanate nanoparticles; these were removed using the cryogenic route. Importantly, the formation of a calcium and phosphorus rich coating around the barium titanate nanoparticles was retained under cryo-conditions, highlighting that significant interactions do occur between nanomaterials and biological media.

Impact of CryoSat-2 for marine gravity field - globally and in the Arctic Ocean

DEFF Research Database (Denmark)

Andersen, Ole Baltazar; Stenseng, Lars; Knudsen, Per

GDR data, NOAA LRM data, but also Level1b (LRM, SAR and SAR-in waveforms) data have been analyzed. A suite of eight different empirical retrackers have been developed and investigated for their ability to predict marine gravity in the Arctic Ocean. The impact of the various improvement offered by Cryo...... days repeat offered by CryoSat-2 provides denser coverage than older geodetic mission data set like ERS-1. Thirdly, the 92 degree inclination of CryoSat-2 is designed to map more of the Arctic Ocean than previous altimetric satellites. Finally, CryoSat-2 is able to operate in two new modes (SAR and SAR......Sat-2 in comparison with conventional satellite altimetry have been studied and quantified both globally but particularly for the Arctic Ocean using a large number of marine and airborne surveys providing “ground truth” marine gravity....

Eye aberration analysis with Zernike polynomials

Science.gov (United States)

Molebny, Vasyl V.; Chyzh, Igor H.; Sokurenko, Vyacheslav M.; Pallikaris, Ioannis G.; Naoumidis, Leonidas P.

1998-06-01

New horizons for accurate photorefractive sight correction, afforded by novel flying spot technologies, require adequate measurements of photorefractive properties of an eye. Proposed techniques of eye refraction mapping present results of measurements for finite number of points of eye aperture, requiring to approximate these data by 3D surface. A technique of wave front approximation with Zernike polynomials is described, using optimization of the number of polynomial coefficients. Criterion of optimization is the nearest proximity of the resulted continuous surface to the values calculated for given discrete points. Methodology includes statistical evaluation of minimal root mean square deviation (RMSD) of transverse aberrations, in particular, varying consecutively the values of maximal coefficient indices of Zernike polynomials, recalculating the coefficients, and computing the value of RMSD. Optimization is finished at minimal value of RMSD. Formulas are given for computing ametropia, size of the spot of light on retina, caused by spherical aberration, coma, and astigmatism. Results are illustrated by experimental data, that could be of interest for other applications, where detailed evaluation of eye parameters is needed.

The effect of cryo-storage on the beta 2-adrenoceptor density and responsiveness in intact human lymphocytes

DEFF Research Database (Denmark)

Ahlquist, P; Johansen, Torben; Friis, U G

1994-01-01

This study evaluates the effect of cryo-storage on beta 2-adrenoceptor number and formation of adenosine 3':5'-cyclic monophosphate (cAMP) in intact human lymphocytes as a measure of the beta 2-adrenoceptor responsiveness. Cryo-storage at -196 degrees C up to 12 months caused no significant......), but changed significantly after long-term storage (3-12 months). We can conclude that lymphocytes can be stored for months for later determination of beta-adrenoceptors. The cryo-storage method described in this paper are, however, only useful for measurements of very large changes in cAMP formation, and our...... results indicate that the method should be further modified in order to preserve the lymphocyte responsiveness after cryo-storage....

Imaging system for creating 3D block-face cryo-images of whole mice

Science.gov (United States)

Roy, Debashish; Breen, Michael; Salvado, Olivier; Heinzel, Meredith; McKinley, Eliot; Wilson, David

2006-03-01

We developed a cryomicrotome/imaging system that provides high resolution, high sensitivity block-face images of whole mice or excised organs, and applied it to a variety of biological applications. With this cryo-imaging system, we sectioned cryo-preserved tissues at 2-40 μm thickness and acquired high resolution brightfield and fluorescence images with microscopic in-plane resolution (as good as 1.2 μm). Brightfield images of normal and pathological anatomy show exquisite detail, especially in the abdominal cavity. Multi-planar reformatting and 3D renderings allow one to interrogate 3D structures. In this report, we present brightfield images of mouse anatomy, as well as 3D renderings of organs. For BPK mice model of polycystic kidney disease, we compared brightfield cryo-images and kidney volumes to MRI. The color images provided greater contrast and resolution of cysts as compared to in vivo MRI. We note that color cryo-images are closer to what a researcher sees in dissection, making it easier for them to interpret image data. The combination of field of view, depth of field, ultra high resolution and color/fluorescence contrast enables cryo-image volumes to provide details that cannot be found through in vivo imaging or other ex vivo optical imaging approaches. We believe that this novel imaging system will have applications that include identification of mouse phenotypes, characterization of diseases like blood vessel disease, kidney disease, and cancer, assessment of drug and gene therapy delivery and efficacy and validation of other imaging modalities.

Possible mechanisms of chromosome aberrations. 2. Formation of aberrations after UV-irradiation

International Nuclear Information System (INIS)

Lebedeva, L.I.

1982-01-01

One of mechanisms of chromosome aberrations after UV-radiation of animal cells initiated by thymine dimerization from different dna threads (by cross joints) and finished in mitosis metaphase is discussed. The model of aberration formation, taking a count of peculiarities of chromosome ansate structure and predicting the important role of chromosome isolation during mitosis in realization of structural aberrations, is suggested. An attempt to present aberration formation under conditions of exact repair is the distinguishing feature of the model

Evaluating the pitch bias of CryoSat exploiting stacks of single look ehoes

Science.gov (United States)

Scagliola, Michele; Tagliani, Nicolas; Fornari, Marco; Bouzinac, Catherine; Parrinello, Tommaso

2014-05-01

CryoSat was launched on the 8th April 2010 and it is the first European ice mission dedicated to monitoring precise changes in the thickness of polar ice sheets and floating sea ice over a 3-year period. CryoSat carries an innovative radar altimeter called the Synthetic Aperture Interferometric Altimeter (SIRAL), that transmits pulses at a high pulse repetition frequency thus making the received echoes phase coherent and suitable for azimuth processing. The attitude information of the spacecraft is provided by star trackers, that have an internal accuracy of few arc-seconds. By analysis of the CryoSat products, two different studies [1, 2] verified the existence of a bias between the pitch reported by the star trackers and the actual pitch of CryoSat during its flight. However those studies, that use two different methods to evaluate the actual pitch, provided different values for the pitch bias. This poster is aimed at describing a further method to estimated the pitch with which the satellite is actually flying by analysis of the stacks of the single look echoes that are accumulated for a given location of sea surface during the Level1 processing. In fact, over ocean the power of the single look echoes for a given point is shaped by the along-track antenna pattern. As a consequence, estimating the angular direction of pointing of the antenna from the stack, an estimate of the pitch can be obtained. Finally, the bias evaluated starting from the pitch measured with the proposed method is compared with the pitch bias measured in [1, 2]. [1] Galin,N. and Wingham, D., Estimating Pitch Angle of CryoSat-2 using the Power Distribution of the Synthetic Aperture, presented at SAR Altimetry Expert Group Meeting, Southampton UK, June 25-27, 2013. [2] Smith, W.H.F. and Scharroo, R., Retracking range, SWH, sigma-naught, and attitude in CryoSat conventional ocean data. In proceedings of Ocean Surface Topography Science Team Meeting. San Diego, October 19-21, 2011.

Statistical Methods for Single-Particle Electron Cryomicroscopy

DEFF Research Database (Denmark)

Jensen, Katrine Hommelhoff

Electron cryomicroscopy (cryo-EM) is a form of transmission electron microscopy, aimed at reconstructing the 3D structure of a macromolecular complex from a large set of 2D projection images, as they exhibit a very low signal-to-noise ratio (SNR). In the single-particle reconstruction (SPR) probl...

CryoSat Ice Processor: High-Level Overview of Baseline-C Data and Quality-Control