title: A negative stain for electron microscopic tomography.
abstract: While negative staining can provide detailed, two-dimensional images of biological structures, the potential of combining tomography with negative staining to provide three-dimensional views has yet to be fully realized. Basic requirements of a negative stain for tomography are that the density and atomic number of the stain are optimal, and that the stain does not degrade or rearrange with the intensive electron dose (~10⁶ e/nm²) needed to collect a full set of tomographic images. A commercially available, tungsten-based stain appears to satisfy these prerequisites. Comparison of the surface structure of negatively stained influenza A virus with previous structural results served to evaluate this negative stain. The combination of many projections of the same structure yielded detailed images of single proteins on the viral surface. Corresponding surface renderings are a good fit to images of the viral surface derived from cryomicroscopy as well as to the shapes of crystallized surface proteins. Negative stain tomography with the appropriate stain yields detailed images of individual molecules in their normal setting on the surface of the influenza A virus.
authors.affiliation: Laboratory of Neurobiology, National Institute of Neurological Disorders and Stroke, Bethesda, MD 20892, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Fera
authors.forename: Jane E
authors.initials: JE
authors.lastname: Farrington
authors.forename: Joshua
authors.initials: J
authors.lastname: Zimmerberg
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Reese
mesh_terms: D004396:Coloring Agents / Q000378:metabolism*; D055032:Electron Microscope Tomography / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D009980:Influenza A virus / Q000648:ultrastructure; D008565:Membrane Proteins / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods*; D014414:Tungsten / Q000378:metabolism*; D014764:Viral Proteins / Q000648:ultrastructure
chemical_list: D004396:Coloring Agents; D008565:Membrane Proteins; D014764:Viral Proteins; D014414:Tungsten