Herpes simplex virus-cell interactions studied by low-fading contrasted immunofluorescence.
The low-fading immunofluorescence with propidium iodide contrast described here is recommended for light and confocal viral antigen identification and other cell biology studies because: (1) it is a simple, rapid, sensitive, and reproducible technique; (2) phase-contrast microscopy is unnecessary; (3) contrast is optimal without blurring the fluorescent labeling; (4) autofluorescence is minimal, even in fixed cells; (5) background staining is minimal; (6) fading is invisible for at least 5-min exposures, even in preparations with weak antigen presentation; (7) fluorescence is stable after storage in the dark at -20 degrees C; (8) fluorochromes are small-sized markers without steric hindrance; and (9) there is no need for silver enhancement or substrate solutions, which increase the risk of diffusion and other artifacts.
Jensen
Helle Lone
HL
Department of Pathology, Rigshospitalet, University Hospital of Copenhagen, Copenhagen, Denmark.
Norrild
Bodil
B
D019411:Clinical Laboratory Techniques; D004847:Epithelial Cells / Q000502:physiology* / Q000821:virology; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D008168:Lung / Q000821:virology; D018139:Simplexvirus / Q000502:physiology*