Journal
JOURNAL OF STRUCTURAL BIOLOGY
Volume 168, Issue 2, Pages 234-239Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2009.07.009
Keywords
Vaccinia virus; Virion structure; Membranes; X-ray microscopy; Cryo-tomography; Electron microscopy; Three-dimensional tomographic reconstruction
Funding
- Foundation Botin
- BESSY-EC-IA-SFS
- German Federal Ministry of Education and Research (BMBF) [05KS4BY1/7]
- Human Frontier Science Program (HFSP) Research Grant [RGP0053/2005-C]
- EU [RII3-CT-2004-506008]
- [BFU2008-2328/BMC]
- [LSHG-CT-2004-502828]
- [CT-2006-037536]
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Vitrified unstained purified vaccinia virus particles have been used as a test sample to evaluate the capabilities of cryo-X-ray tomography. Embedded in a thick layer of vitreous ice, the viral particles representing the mature form of the virus (MV) were visualized using full-field transmission X-ray tomography. The tomographic reconstructions reveal the viral brick-shaped characteristic structures with a size of 250 x 270 x 360 nm(3). The X-ray tomograms show the presence of a clearly defined external envelope, together with an inner core surrounded by an internal envelope, including areas with clear differential density, which correlate well with those features previously described for these viral particles using electron microscopy analyses. A quantitative assessment of the resolution attained in X-ray and electron tomograms of the viral particles prepared under the same conditions yields values of 25.7 and 6.7 nm half-pitch, respectively. Although the resolution of the X-ray microscope is well above the dimensions of the membranous compartments, the strong differential contrast exhibited makes it possible to precisely reveal them without any contrasting reagent within this small and complex biological sample. (C) 2009 Elsevier Inc. All rights reserved.
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