Rapid virus subunit visualization by direct sedimentation of samples on electron microscope grids.

Abstract

Airfuge direct ultracentrifugation of viral samples on electron microscope grids offers a rapid way for concentrating viral particles or subunits to facilitate their detection and study. Using the A-100 fixed angle rotor (30 degrees) with a K factor of 19 at maximum speed (95,000 rpm), samples up to 240 microliters can be prepared for electron microscopy observation in a few minutes: observation time is decreased and structural details are highlighted. Using latex spheres to calculate the increase in sensitivity compared to the inverted drop procedure, we obtained a 10- to 40-fold increase in sensitivity depending on the size of particles. Application of this technique to rubella virus permitted better visualization of viral membrane subunits on the particles. Rubella hemagglutinin immuno-stimulating complexes preparations were also better visualized and their morphology conserved after direct ultracentrifugation on the specimen grids. Similar observations are reported for respiratory syncytial virus associated subunits.

• Authors: R Alain, F Nadon, C Séguin, P Payment, M Trudel
• Journal: Journal of virological methods (J Virol Methods) Vol. 16 Issue 3 Pg. 209-16 (Jun 1987) ISSN: 0166-0934 [Print] Netherlands
• PMID: 3654898 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Topics

• Animals
• Cell Line
• Microscopy, Electron
• Microspheres
• Respiratory Syncytial Viruses (ultrastructure)
• Rubella virus (ultrastructure)
• Ultracentrifugation (methods)
• Vero Cells