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

• Main Authors: Demeestere Lien, Mast Jan
• Format: Article
• Language: English
• Published: BMC 2009-02-01
• Series: Diagnostic Pathology
• Online Access: http://www.diagnosticpathology.org/content/4/1/5

<p>Abstract</p> <p>Background</p> <p>Electron tomographic analysis can be combined with the simple and rapid negative staining technique used in electron microscopy based virus diagnosis.</p> <p>Methods</p> <p>Standard negative staining of representative examples of parapoxviruses and paramyxoviruses was combined with electron tomographic analysis.</p> <p>Results</p> <p>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.</p> <p>Conclusion</p> <p>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.</p>