Electron Microscopy for Rapid Diagnosis of Emerging Infectious Agents1
Figure 5. Comparison of herpesvirus appearance after positive and negative stain electron microscopic. A. Positive staining. Samples undergo a lengthy process of fixation, incubation with heavy metal ions (osmium, uranyl), dehydration, embedment, ultrathin sectioning, and staining. Chemical moieties in the object show differential affinities for the heavy metal stains, resulting in a clear outline of the viral bilayer envelope, viral envelope proteins, nucleocapsid, and the dense nucleic acid containing core. B. Negative staining. After a brief fixation, samples are mounted directly on electron microscopic grids and stained as in Figure 4. The electron-dense stain (phosphotungstic acid [phosphotungstic acid], uranyl acetate, and the like) penetrates the virion and embeds the particle in a matrix of stain. Due to density differences between the stain and weakly scattering biological components of the virion, the virion appears as a transparent and detailed reverse (negative) image. Penetration of stain into the nucleocapsid provides a dense core with the crenellated appearance presented by the central channel of capsomers on the nucleocapsid surface. Viral surface proteins appear as projections from the labile envelope. phosphotungstic acid. Bar = 100 nm.