scholarly journals Capturing Enveloped Viruses on Affinity Grids for Downstream Cryo-Electron Microscopy Applications

2013 ◽  
Vol 20 (1) ◽  
pp. 164-174 ◽  
Author(s):  
Gabriella Kiss ◽  
Xuemin Chen ◽  
Melinda A. Brindley ◽  
Patricia Campbell ◽  
Claudio L. Afonso ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Measles Virus ◽  
Influenza A ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Nitrilotriacetic Acid ◽  
Dimensional Structure ◽  
Enveloped Viruses ◽  
Cryo Electron Microscopy ◽  
Human Immunodeficiency Virus Virus

AbstractElectron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane-derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids (AG) for use in both conventional EM and cryo-EM/ET applications. We examined the utility of AG for the selective capture of human immunodeficiency virus virus-like particles, influenza A, and measles virus. We applied nickel-nitrilotriacetic acid lipid layers in combination with molecular adaptors to selectively adhere the viruses to the AG surface. This further development of the AG method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analyses.

Three-Dimensional Structure of Helical and Zigzagged Nanowires Using Electron Tomography

2008 ◽  
Vol 1144 ◽  
Author(s):  
Han Sung Kim ◽  
Yoon Myung ◽  
Chang Hyun Kim ◽  
Seung Yong Bae ◽  
Jae-Pyoung Ahn ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
High Resolution ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Helical Structure ◽  
Axial Direction ◽  
Dimensional Structure ◽  
Gan Nanowires ◽  
Transmission Electron

ABSTRACTElectron tomography and high-resolution transmission electron microscopy were used to characterize the unique three-dimensional structures of helical or zigzagged GaN, ZnGa2O4 and Zn2SnO4 nanowires. The helical GaN nanowires adopt a helical structure that consists of six equivalent <0-111> growth directions with the axial [0001] direction. The ZnGa2O4 nanosprings have four equivalent <011> growth directions with the [001] axial direction. The zigzagged Zn2SnO4 nanowires consisted of linked rhombohedrons structure having the side edges matched to the <011> direction, and the [111] axial direction.

Cryo-Electron Microscopy of Aura Viruses

1998 ◽  
Vol 4 (S2) ◽  
pp. 946-947
Author(s):  
W. Zhang ◽  
N. H. Olson ◽  
B. R. McKinney ◽  
R. J. Kuhn ◽  
T. S. Baker
Keyword(s):  
Electron Microscopy ◽  
Three Dimensional ◽  
Single Copy ◽  
Wild Type Virus ◽  
Genomic Rna ◽  
Wild Type ◽  
Subgenomic Rna ◽  
Enveloped Viruses ◽  
Virus Particles ◽  
Cryo Electron Microscopy

Alphaviruses are a group of enveloped viruses in the Togaviridae family. Studies of several alphaviruses, including Ross River, Sindbis and Semliki Forest viruses, by cryo-electron microscopy (cryo-EM), three-dimensional (3D) image resconstruction and other techniques have illustrated that these spherical viruses have a T=4, multi-layered structure.Aura virus, which is closely related to Sindbis, was first isolated in South America. Unlike the other alphaviruses, both genomic RNA (12kb, 49S) and subgenomic RNA(4.2kb, 26S) are encapsidated efficiently and form mature virions. Studies on negatively-stained virus particles demonstrated that there are two major size classes. The first contains particles of ∼72nm diameter, which are most similar to wild type virus, whereas the second class includes particles of ∼62nm in diameter. The 72nm particles are believed to have one copy of genomic RNA or one to three copies of subgenomic RNA, and a T=4 structure. The 62nm particles probably only have a single copy of subgenomic RNA and are presumed to be T=3 structures.

Cryo-Electron Microscopy of Spiroplasma Virus SpV4

1997 ◽  
Vol 3 (S2) ◽  
pp. 87-88
Author(s):  
P.R. Chipman ◽  
R. Mckenna ◽  
J. Renaudin ◽  
T.S. Baker
Keyword(s):  
Electron Microscopy ◽  
Cross Section ◽  
Outer Surface ◽  
Average Distance ◽  
Three Dimensional ◽  
Longitudinal Axis ◽  
Lytic Cycle ◽  
Dimensional Structure ◽  
Circular Dna ◽  
Cryo Electron Microscopy

Spiroplasma, a wall-free prokaryote of the class Mollicutes, is host to a small, naked, single-stranded DNA, isometric virus. Spiroplasma virus SpV4 belongs to the Microviridae family, members of which are non-enveloped, have icosahedral capsids, release progeny through a lytic cycle, and contain circular DNA.Measurements obtained from negatively stained SpV4 particles revealed a nucleocapsid of 27nm in diameter (figure 1). The three-dimensional structure reported here, obtained from unstained particles suspended in a layer of vitreous ice (figure 2), is in agreement with these earlier results, suggesting a 27nm average distance through the nucleocapsid (figure 3). Unreported in earlier studies is the presence of a 6nm, mushroom-shaped protrusion (made up of a stalk, 2.3nm long and 1.3nm wide, and a globular bud of dimensions ≈4.0×4.0×3.7nm) stemming from an ≈1.5nm deep depression at each of the 3-fold icosahedral axes of the virion. A cross section through the longitudinal axis of one protuberance (figure 4) reveals a cylindrical dimple (≈1.0nm in diameter and 2.3nm deep), originating on the axis of the outer surface of the globular bud domain.

scholarly journals Three dimensional structure of the anthrax toxin translocon-lethal factor complex by cryo-electron microscopy

2013 ◽  
Vol 22 (5) ◽  
pp. 586-594 ◽  
Author(s):  
E. P. Gogol ◽  
N. Akkaladevi ◽  
L. Szerszen ◽  
S. Mukherjee ◽  
L. Chollet-Hinton ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Lethal Factor ◽  
Three Dimensional ◽  
Anthrax Toxin ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
Cryo Electron Microscopy
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