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Diagn Pathol. 2009 Feb 10;4:5. doi: 10.1186/1746-1596-4-5.

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

Author information

1
EM-unit, CODA-CERVA, Groeselenberg 99, Brussels, Belgium. jamas@var.fgov.be

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.

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