protocol_161.txt

Electron microscopy for virus identification and virus assemblage characterization
5µL of an enrichment culture, or 0.2 µm filtered and centrifuged (10 min at 3000g) cell-free supernatant, is spotted onto carbon/formvar-coated electron microscope grids (Ted Pella or EM Sciences).
It is then allowed to absorb for 2 min.
Remove sample from grid by slowly bringing a small (ca. 1cm2) piece of filter paper perpendicular to the grid to the side of the grid.
The sample will be removed by wicking.
Stain grid by placing on a 5μL drop of 2% Uranyl Acetate (or Phosphotungstate) for 15-30 seconds.
Remove stain by wicking as above.
Air dry for at least 10 minutes.
Samples are examined by transmission electron microscopy (TEM), e.g., JEOL 100 cx, operated at 100 keV.