scholarly journals Reconstruction of Average Subtracted Tubular Regions (RASTR) Enables Structure Determination of Tubular Filaments by Cryo-EM

2019 ◽  
Author(s):  
Peter S. Randolph ◽  
Scott M. Stagg
Keyword(s):  
Electron Microscopy ◽  
Processing Technique ◽  
Processing Method ◽  
Tubular Structures ◽  
Global Order ◽  
Helical Symmetry ◽  
Cellular Processes ◽  
New Processing ◽  
Helical Parameters

AbstractAs the field of electron microscopy moves forward, the increasing complexity of samples being produced demand more involved processing methods. In this study, we have developed a new processing method for generating 3D reconstructions of tubular structures. Tubular biomolecules are common throughout many cellular processes and are appealing targets for a variety of biophysical research. Processing of tubules with helical symmetry is relatively straightforward for electron microscopy if the helical parameters are known, but tubular structures that deviate from helical symmetry (asymmetrical components, local but no global order, etc) present myriad issues. Here we present a new processing technique called Reconstruction of Average Subtracted Tubular Regions (RASTR), which was developed to reconstruct tubular structures without applying symmetry. We explain the RASTR approach and quantify its performance using three examples: a simulated symmetrical tubular filament, a symmetrical tubular filament from cryo-EM data, and a membrane tubule coated with locally ordered but not globally ordered proteins.

A New Image Processing Technique for STEM

1974 ◽  
Vol 32 ◽  
pp. 432-433
Author(s):  
Yasushi Kokubo ◽  
Hirotami Koike ◽  
Teruo Someya
Keyword(s):  
Electron Microscopy ◽  
Image Processing ◽  
Scanning Electron Microscopy ◽  
Elastic Scattering ◽  
Field Emission ◽  
Processing Technique ◽  
Image Processing Technique ◽  
Energy Analyzer ◽  
Scanning Microscope ◽  
Scanning Electron

One of the advantages of scanning electron microscopy is the capability for processing the image contrast, i.e., the image processing technique. Crewe et al were the first to apply this technique to a field emission scanning microscope and show images of individual atoms. They obtained a contrast which depended exclusively on the atomic numbers of specimen elements (Zcontrast), by displaying the images treated with the intensity ratio of elastically scattered to inelastically scattered electrons. The elastic scattering electrons were extracted by a solid detector and inelastic scattering electrons by an energy analyzer. We noted, however, that there is a possibility of the same contrast being obtained only by using an annular-type solid detector consisting of multiple concentric detector elements.

Complex Vesicles, Microtubules, and Cytoplasmic Filaments in the Endothelial Cells of Normal Dog Aorta

1968 ◽  
Vol 26 ◽  
pp. 172-173
Author(s):  
C. N. Sun ◽  
J. J. Ghidoni
Keyword(s):  
Electron Microscopy ◽  
Endothelial Cells ◽  
Cross Sections ◽  
Functional Significance ◽  
Tubular Structures ◽  
Aldehyde Fixation ◽  
Cytoplasmic Filaments

Endothelial cells in longitudinal and cross sections of aortas from 3 randomly selected “normal” mongrel dogs were studied by electron microscopy. Segments of aorta were distended with cold cacodylate buffered 5% glutaraldehyde for 10 minutes prior to being cut into small, well oriented tissue blocks. After an additional 1-1/2 hour period in glutaraldehyde, the tissue blocks were well rinsed in buffer and post-fixed in OsO4. After dehydration they were embedded in a mixture of Maraglas, D.E.R. 732, and DDSA.Aldehyde fixation preserves the filamentous and tubular structures (300 Å and less) for adequate demonstration and study. The functional significance of filaments and microtubules has been recently discussed by Buckley and Porter; the precise roles of these cytoplasmic components remains problematic. Endothelial cells in canine aortas contained an abundance of both types of structures.

Enhanced information from biological materials through technological improvements in cryo-electron microscopy

1992 ◽  
Vol 50 (1) ◽  
pp. 788-789
Author(s):  
Marc J.C. de Jong ◽  
Wim M. Busing ◽  
Max T. Otten
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
Biological Materials ◽  
Electron Crystallography ◽  
Cryo Electron Microscopy ◽  
Transmission Electron ◽  
The Many ◽  
Technological Improvements ◽  
Beam Damage

Biological materials damage rapidly in the electron beam, limiting the amount of information that can be obtained in the transmission electron microscope. The discovery that observation at cryo temperatures strongly reduces beam damage (in addition to making it unnecessaiy to use chemical fixatives, dehydration agents and stains, which introduce artefacts) has given an important step forward to preserving the ‘live’ situation and makes it possible to study the relation between function, chemical composition and morphology.Among the many cryo-applications, the most challenging is perhaps the determination of the atomic structure. Henderson and co-workers were able to determine the structure of the purple membrane by electron crystallography, providing an understanding of the membrane's working as a proton pump. As far as understood at present, the main stumbling block in achieving high resolution appears to be a random movement of atoms or molecules in the specimen within a fraction of a second after exposure to the electron beam, which destroys the highest-resolution detail sought.

scholarly journals Determination of intermediate state structures in the opening pathway of SARS-CoV-2 spike using cryo-electron microscopy

2021 ◽  
Author(s):  
Z. Faidon Brotzakis ◽  
Thomas Lohr ◽  
Michele Vendruscolo
Keyword(s):  
Infectious Disease ◽  
Electron Microscopy ◽  
Severe Acute Respiratory Syndrome ◽  
Intermediate State ◽  
Therapeutic Interventions ◽  
Cryo Electron Microscopy ◽  
Welfare Systems

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes COVID-19, a highly infectious disease that is severely affecting our society and welfare systems. In order to develop therapeutic interventions against this...

scholarly journals A Magnetic Nanocomposite Modifier for Improved Ultrasensitive Detection of Hexavalent Chromium in Water Samples

Chemosensors ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 189
Author(s):  
Nuša Hojnik ◽  
Olivija Plohl ◽  
Matjaž Finšgar
Keyword(s):  
Electron Microscopy ◽  
Analytical Performance ◽  
Magnetic Nanocomposite ◽  
Ex Situ ◽  
Trace Determination ◽  
Silica Layer ◽  
X Ray ◽  
Square Wave ◽  
Static Contact

In this work, different electrodes were employed for the determination of Cr(VI) by the cathodic square‑wave voltammetry (SWV) technique and the square-wave adsorptive stripping voltammetry (SWAdSV) technique in combination with diethylenetriaminepentaacetic acid. Using SWV, a comparison of the analytical performance of the bare glassy carbon electrode (GCE), ex situ electrodes (antimony-film—SbFE, copper-film—CuFE, and bismuth-film—BiFE), and the GCE modified with a new magnetic nanocomposite (MNC) material was performed. First, the MNC material was synthesized, i.e., MNPs@SiO2@Lys, where MNPs stands for magnetic maghemite nanoparticles, coated with a thin amorphous silica (SiO2) layer, which was additionally functionalized with derived lysine (Lys). The crystal structure of the prepared MNCs was confirmed by X-ray powder diffraction (XRD), while the morphology and nano-size of the MNCs were investigated by field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM), where TEM was additionally used to observe the MNP core and silica layer thickness. The presence of functional groups of the MNCs was investigated by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and surface analysis was performed by X-ray photoelectron spectroscopy (XPS). The hydrophilicity of the modified electrodes was also tested by static contact angle measurements. Then, MNPs@SiO2@Lys was applied onto the electrodes and used with the SWV and SWAdSV techniques. All electrodes tested with the SWV technique were effective for Cr(VI) trace determination. On the other hand, the SWAdSV technique was required for ultra-trace determination of Cr(VI). Using the SWAdSV technique, it was shown that a combination of ex situ BiFE with the deposited MNPs@SiO2@Lys resulted in excellent analytical performance (LOQ = 0.1 µg/L, a linear concentration range of 0.2–2.0 µg/L, significantly higher sensitivity compared to the SWV technique, an RSD representing reproducibility of 9.0%, and an average recovery of 98.5%). The applicability of the latter system was also demonstrated for the analysis of a real sample.

scholarly journals Determination of the 3-D Magnetic Vector Potential using Lorentz Transmission Electron Microscopy

2009 ◽  
Vol 15 (S2) ◽  
pp. 134-135 ◽  
Author(s):  
C Phatak ◽  
E Humphrey ◽  
M DeGraef ◽  
A Petford-Long
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Vector Potential ◽  
Magnetic Vector Potential ◽  
Magnetic Vector ◽  
Transmission Electron ◽  
Lorentz Transmission Electron Microscopy

Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 – July 30, 2009

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