scholarly journals In-depth structural analysis of glycans in the gas phase

2019 ◽  
Vol 10 (5) ◽  
pp. 1272-1284 ◽  
Author(s):  
Eike Mucha ◽  
Alexandra Stuckmann ◽  
Mateusz Marianski ◽  
Weston B. Struwe ◽  
Gerard Meijer ◽  
...  
Keyword(s):  
Structural Analysis ◽  
High Resolution ◽  
Gas Phase ◽  
High Throughput ◽  
Glycan Analysis ◽  
The Past

Although there have been substantial improvements in glycan analysis over the past decade, the lack of both high-resolution and high-throughput methods hampers progress in glycomics.

scholarly journals Epoxidized graphene grid for high-throughput high-resolution cryoEM structural analysis

2021 ◽  
Author(s):  
Junso Fujita ◽  
Fumiaki Makino ◽  
Haruyasu Asahara ◽  
Maiko Moriguchi ◽  
Shota Kumano ◽  
...  
Keyword(s):  
Structural Analysis ◽  
High Resolution ◽  
Data Collection ◽  
High Throughput ◽  
Particle Density ◽  
Orientation Distribution ◽  
Biological Macromolecules ◽  
Density Maps ◽  
V1 Atpase ◽  
Density Map

Many specimens suffer from low particle density and/or preferred orientation in cryoEM specimen grid preparation, making data collection and structure determination time consuming. We developed an epoxidized graphene grid (EG-grid) that effectively immobilizes protein particles by applying an oxidation reaction using photoactivated ClO2· and further chemical modification. The particle density and orientation distribution are both dramatically improved, having enabled us to reconstruct the density map of GroEL and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), at 1.99 and 2.16 Å resolution from only 504 and 241 micrographs, respectively. A low concentration sample solution of 0.1 mg ml–1 was sufficient to reconstruct a 3.10 Å resolution density map of SARS-CoV-2 spike protein from 1,163 micrographs. The density maps of V1-ATPase, β-galactosidase, and apoferritin were also reconstructed at 3.03, 1.81, and 1.29 Å resolution, respectively. These results indicate that the EG-grid will be a powerful tool for high-throughput cryoEM data collection to accelerate high-resolution structural analysis of biological macromolecules.

Structural analysis of the surface-layer protein of spirillum serpens by high-resolution electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 738-739
Author(s):  
W. H. Wu ◽  
R. M. Glaeser
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Structural Analysis ◽  
High Resolution ◽  
Low Dose ◽  
High Resolution Electron Microscopy ◽  
Optical Diffraction ◽  
Surface Layer Protein ◽  
Morphological Unit ◽  
Resolution Electron

Spirillum serpens possesses a surface layer protein which exhibits a regular hexagonal packing of the morphological subunits. A morphological model of the structure of the protein has been proposed at a resolution of about 25 Å, in which the morphological unit might be described as having the appearance of a flared-out, hollow cylinder with six ÅspokesÅ at the flared end. In order to understand the detailed association of the macromolecules, it is necessary to do a high resolution structural analysis. Large, single layered arrays of the surface layer protein have been obtained for this purpose by means of extensive heating in high CaCl2, a procedure derived from that of Buckmire and Murray. Low dose, low temperature electron microscopy has been applied to the large arrays.As a first step, the samples were negatively stained with neutralized phosphotungstic acid, and the specimens were imaged at 40,000 magnification by use of a high resolution cold stage on a JE0L 100B. Low dose images were recorded with exposures of 7-9 electrons/Å2. The micrographs obtained (Fig. 1) were examined by use of optical diffraction (Fig. 2) to tell what areas were especially well ordered.

RNA polymerase: Preparation and structural analysis of helical polymers

1984 ◽  
Vol 42 ◽  
pp. 152-153
Author(s):  
E. Loren Buhle ◽  
Pamela Rew ◽  
Ueli Aebi
Keyword(s):  
Structural Analysis ◽  
Rna Polymerase ◽  
Molecular Level ◽  
X Ray Diffraction ◽  
Helical Polymers ◽  
X Ray ◽  
E Coli ◽  
The Past ◽  
Ordered Arrays ◽  
Low Ionic Strength

While DNA-dependent RNA polymerase represents one of the key enzymes involved in transcription and ultimately in gene expression in procaryotic and eucaryotic cells, little progress has been made towards elucidation of its 3-D structure at the molecular level over the past few years. This is mainly because to date no 3-D crystals suitable for X-ray diffraction analysis have been obtained with this rather large (MW ~500 kd) multi-subunit (α2ββ'ζ). As an alternative, we have been trying to form ordered arrays of RNA polymerase from E. coli suitable for structural analysis in the electron microscope combined with image processing. Here we report about helical polymers induced from holoenzyme (α2ββ'ζ) at low ionic strength with 5-7 mM MnCl2 (see Fig. 1a). The presence of the ζ-subunit (MW 86 kd) is required to form these polymers, since the core enzyme (α2ββ') does fail to assemble into such structures under these conditions.

The Contribution of Intermediate-Voltage, High-Resolution Electron Microscopy In Materials Science: An Appraisal

1990 ◽  
Vol 48 (1) ◽  
pp. 478-479
Author(s):  
John L. Hutchison
Keyword(s):  
High Resolution ◽  
Materials Science ◽  
High Resolution Electron Microscopy ◽  
Optical Diffraction ◽  
The Past ◽  
Resolution Electron ◽  
Extreme Sensitivity ◽  
Electron Microscopes ◽  
New Generation ◽  
Small Metal Particles

Over the past five years or so the development of a new generation of high resolution electron microscopes operating routinely in the 300-400 kilovolt range has produced a dramatic increase in resolution, to around 1.6 Å for “structure resolution” and approaching 1.2 Å for information limits. With a large number of such instruments now in operation it is timely to assess their impact in the various areas of materials science where they are now being used. Are they falling short of the early expectations? Generally, the manufacturers’ claims regarding resolution are being met, but one unexpected factor which has emerged is the extreme sensitivity of these instruments to both floor-borne and acoustic vibrations. Successful measures to counteract these disturbances may require the use of special anti-vibration blocks, or even simple oil-filled dampers together with springs, with heavy curtaining around the microscope room to reduce noise levels. In assessing performance levels, optical diffraction analysis is becoming the accepted method, with rotational averaging useful for obtaining a good measure of information limits. It is worth noting here that microscope alignment becomes very critical for the highest resolution.In attempting an appraisal of the contributions of intermediate voltage HREMs to materials science we will outline a few of the areas where they are most widely used. These include semiconductors, oxides, and small metal particles, in addition to metals and minerals.

The Contribution of Phonon-Scattered Electrons to High-Resolution Electron Micrographs of Crystals

1990 ◽  
Vol 48 (1) ◽  
pp. 62-63
Author(s):  
H. Kohl
Keyword(s):  
High Resolution ◽  
Spherical Aberration ◽  
High Resolution Electron Microscopy ◽  
Static Potential ◽  
Qualitative Comparison ◽  
The Past ◽  
Resolution Electron ◽  
High Resolution Images ◽  
High Resolution Electron Micrographs ◽  
Extract Information

High-Resolution Electron Microscopy is able to determine structures of crystals and interfaces with a spatial resolution of somewhat less than 2 Å. As the image is strongly dependent on instrumental parameters, notably the defocus and the spherical aberration, the interpretation of micrographs necessitates a comparison with calculated images. Whereas one has often been content with a qualitative comparison of theory with experiment in the past, one is currently striving for quantitative procedures to extract information from the images [1,2]. For the calculations one starts by assuming a static potential, thus neglecting inelastic scattering processes.We shall confine the discussion to periodic specimens. All electrons, which have only been elastically scattered, are confined to very few directions, the Bragg spots. In-elastically scattered electrons, however, can be found in any direction. Therefore the influence of inelastic processes on the elastically (= Bragg) scattered electrons can be described as an attenuation [3]. For the calculation of high-resolution images this procedure would be correct only if we had an imaging energy filter capable of removing all phonon-scattered electrons. This is not realizable in practice. We are therefore forced to include the contribution of the phonon-scattered electrons.

Cross Section Analysis of Cu Filled TSVs Based on High Throughput Plasma-FIB Milling

2012 ◽  
Author(s):  
Frank Altmann ◽  
Jens Beyersdorfer ◽  
Jan Schischka ◽  
Michael Krause ◽  
German Franz ◽  
...  
Keyword(s):  
High Resolution ◽  
Cross Section ◽  
High Throughput ◽  
Cross Sections ◽  
Electron Backscatter Diffraction ◽  
Grain Structure ◽  
Electron Backscatter ◽  
Section Surface ◽  
Backscatter Diffraction ◽  
Section Analysis

Abstract In this paper the new Vion™ Plasma-FIB system, developed by FEI, is evaluated for cross sectioning of Cu filled Through Silicon Via (TSV) interconnects. The aim of the study presented in this paper is to evaluate and optimise different Plasma-FIB (P-FIB) milling strategies in terms of performance and cross section surface quality. The sufficient preservation of microstructures within cross sections is crucial for subsequent Electron Backscatter Diffraction (EBSD) grain structure analyses and a high resolution interface characterisation by TEM.

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