Serial Block-Face Imaging and its Potential for Reconstructing Diminutive Cell Systems: A Case Study from Arthropods

2014 ◽  
Vol 20 (3) ◽  
pp. 946-955 ◽  
Author(s):  
Elisabeth Lipke ◽  
Thomas Hörnschemeyer ◽  
Anahita Pakzad ◽  
Christopher R. Booth ◽  
Peter Michalik
Keyword(s):  
Electron Microscopy ◽  
Three Dimensional ◽  
Ultrastructural Level ◽  
Cellular Level ◽  
Cellular Systems ◽  
Transmission Electron ◽  
High Lateral Resolution ◽  
Dimensional Reconstruction ◽  
Block Face

AbstractUntil recently, three-dimensional reconstruction on an ultrastructural level was only possible using serial section transmission electron microscopy (ssTEM). However, ssTEM is highly challenging and prone to artifacts as, e.g., section loss and image distortions. New methods, such as serial block-face scanning electron microscopy (SBFSEM) overcome these limitations and promise a high lateral resolution. However, little is known about the usability of SBFSEM in diminutive, but highly complex cellular systems. We used spider sperm (~3 µm in diameter), which fulfills these conditions, to analyze the potential of SBFSEM compared with ssTEM. Our data suggest that the resolution obtained by SBFSEM allows depicting structures on a cellular level and is sufficient to discriminate subcellular components, but is highly dependent on previous staining procedures and electron density of the target structures.

scholarly journals Three-dimensional reconstruction on cell level: case study elucidates the ultrastructure of the spinning apparatus of Embia sp. (Insecta: Embioptera)

2016 ◽  
Vol 3 (10) ◽  
pp. 160563 ◽  
Author(s):  
Sebastian Büsse ◽  
Thomas Hörnschemeyer ◽  
Christian Fischer
Keyword(s):  
High Resolution ◽  
Three Dimensional ◽  
Silk Gland ◽  
Cellular Level ◽  
Ultrastructural Organization ◽  
Three Dimensional Reconstruction ◽  
Dimensional Reconstruction ◽  
Block Face ◽  
Morphological Research

Spinning is a phenomenon not only present in spiders, but also in many other arthropods. The functional morphology and complexity of spinning organs is often poorly understood. Their elements are minute and studying them poses substantial methodological difficulties. This study presents a three-dimensional reconstruction of a silk gland of Embia sp. on cellular level, based on serial sections acquired with serial block-face scanning electron microscopy (SBFSEM) to showcase the power of this method. Previous studies achieved either high resolution to elucidate the ultrastructure or satisfying three-dimensional representations. The high-resolution achieved by SBFSEM can be easily used to reconstruct the three-dimensional ultrastructural organization of cellular structures. The herein investigated spinning apparatus of Embioptera can be taken as an example demonstrating the potential of this method. It was possible to reconstruct a multinucleated silk gland containing 63 nuclei. We focused on the applicability of this method in the field of morphological research and provide a step-by-step guide to the methodology. This will help in applying the method to other arthropod taxa and will help significantly in adapting the method to other animals, animal parts and tissues.

Three-dimensional reconstruction of cell morphology using intermediate voltage electron microscopy and computer graphics

1987 ◽  
Vol 45 ◽  
pp. 590-591 ◽  
Author(s):  
Julian P. Heath ◽  
Ming Hsiu Ho ◽  
Lee D. Peachey
Keyword(s):  
Electron Microscopy ◽  
Three Dimensional ◽  
University Of Pennsylvania ◽  
Stereoscopic Display ◽  
Collagen Gels ◽  
Biomedical Image ◽  
Voltage Electron ◽  
Specimen Holder ◽  
Transmission Electron ◽  
Dimensional Reconstruction

The Intermediate Voltage Electron Microscopy and Biomedical Image Analysis facility at the University of Pennsylvania is a National Resource supported by NIH and provides users with facilities for transmission electron microscopy and digital image processing. The facility comprises a JEOL 4000EX transmission electron microscope with a 360 degree goniometer specimen holder, a VAX 11/750 computer, and a Raster Technologies Inc. Model One/380 Graphics work station with two high resolution 1280x1024 pixel color video display monitors for stereoscopic display. We are using this facility to examine the morphology of fibroblasts migrating through fibrillar collagen gels: these matrices closely model the environments encountered by migratory cells during embryonic morphogenesis.The identification and three dimensional localization of structures in stereoscopic electron micrographs of thick sections of cells can be hampered by diffuse boundaries and by the superposition of details with similar electron density.

scholarly journals Three-dimensional reconstruction of an actin bundle.

1988 ◽  
Vol 107 (2) ◽  
pp. 597-611 ◽  
Author(s):  
E S Bullitt ◽  
D J DeRosier ◽  
L M Coluccio ◽  
L G Tilney
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Three Dimensional ◽  
Oblate Spheroid ◽  
Potassium Thiocyanate ◽  
Dimensional Structure ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
Dimensional Reconstruction ◽  
Molecular Masses

We present the three-dimensional structure of an actin filament bundle from the sperm of Limulus. The bundle is a motile structure which by changing its twist, converts from a coiled to an extended form. The bundle is composed of actin plus two auxiliary proteins of molecular masses 50 and 60 kD. Fraying the bundle with potassium thiocyanate created three classes of filaments: actin, actin plus the 60-kD protein, and actin plus both the auxiliary proteins. We examined these filaments by transmission electron microscopy and scanning transmission electron microscopy (STEM). Three-dimensional reconstructions from electron micrographs allowed us to visualize the actin subunit and the 60- and 50-kD subunits bound to it. The actin subunit appears to be bilobed with dimensions 70 X 40 X 35 A. The inner lobe of the actin subunit, located at 20 A radius, is a prolate ellipsoid, 50 X 25 A; the outer actin lobe, at 30 A radius, is a 35-A-diam spheroid. Attached to the inner lobe of actin is the 60-kD protein, an oblate spheroid, 55 X 40 A, at 50 A radius. The armlike 50-kD protein, at 55 A radius, links the 60-kD protein on one of actin's twin strands to the outer lobe of the actin subunit on the opposite strand. We speculate that the 60-kD protein may be a bundling protein and that the 50-kD protein may be responsible for the change in twist of the filaments which causes extension of the bundle.

The fine structure and organization of the tail musculature of the cercaria of Schistosoma mansoni

Parasitology ◽  
1974 ◽  
Vol 68 (2) ◽  
pp. 147-154 ◽  
Author(s):  
C. J. Nuttman
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Schistosoma Mansoni ◽  
Three Dimensional ◽  
Three Dimensional Reconstruction ◽  
Transmission Electron ◽  
Dimensional Reconstruction ◽  
The Relationship

The fine structure and organization of schistosome tail musculature has been investigated by transmission electron microscopy. A three-dimensional reconstruction is presented and the relationship between morphology and cercarial behaviour is discussed.

scholarly journals Cryo-electron microscopy reconstructions of triatoma virus particles: a clue to unravel genome delivery and capsid disassembly

2013 ◽  
Vol 94 (5) ◽  
pp. 1058-1068 ◽  
Author(s):  
J. Agirre ◽  
G. Goret ◽  
M. LeGoff ◽  
R. Sánchez-Eugenia ◽  
G. A. Marti ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Conformational Changes ◽  
Three Dimensional ◽  
Intact Protein ◽  
Virus Family ◽  
Transmission Electron ◽  
Triatoma Virus ◽  
Dimensional Reconstruction ◽  
Subsequent Step ◽  
Protein Shell

Triatoma virus (TrV) is a member of the insect virus family Dicistroviridae and consists of a small, non-enveloped capsid that encloses its positive-sense ssRNA genome. Using cryo-transmission electron microscopy and three-dimensional reconstruction techniques combined with fitting of the available crystallographic models, this study analysed the capsids corresponding to mature and several RNA-empty TrV particles. After genome release, the resulting reconstruction of the empty capsids displayed no prominent conformational changes with respect to the full virion capsid. The results showed that RNA delivery led to empty capsids with an apparent overall intact protein shell and suggested that, in a subsequent step, empty capsids disassemble into small symmetrical particles. Contrary to what is observed upon genome release in mammalian picornaviruses, the empty TrV capsid maintained a protein shell thickness and size identical to that in full virions.

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