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Open Access Highly Accessed Research

Electron tomography of negatively stained complex viruses: application in their diagnosis

Jan Mast* and Lien Demeestere

Author Affiliations

EM-unit, CODA-CERVA, Groeselenberg 99, Brussels, Belgium

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Diagnostic Pathology 2009, 4:5  doi:10.1186/1746-1596-4-5

Published: 10 February 2009

Abstract

Background

Electron tomographic analysis can be combined with the simple and rapid negative staining technique used in electron microscopy based virus diagnosis.

Methods

Standard negative staining of representative examples of parapoxviruses and paramyxoviruses was combined with electron tomographic analysis.

Results

Digital sectioning of reconstructions of these viruses at a selected height demonstrated the viral ultrastructure in detail, including the characteristic diagnostic features like the surface threads on C-particles of a parapoxvirus and individual glycoproteins and the internal nucleoprotein strand of Newcastle disease virus. For both viruses, deformation and flattening were observed.

Conclusion

The combination of negative staining of complex viruses with electron tomographic analysis, allows visualizing and measuring artifacts typical for negative staining. This approach allows sharp visualisation of structures in a subnanometer-thick plane, avoiding blurring due to superposition which is inherent to TEM. In selected examples, such analyses can improve diagnosis of viral agents.