Vaccinia Virus and Poxvirology

Volume 269 of the series Methods in Molecular Biology pp 205-218

Pox, Dyes, and Videotape

Making Movies of GFP-Labeled Vaccinia Virus
  • Brian M. WardAffiliated withLaboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health

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The large size of poxvirus virions (approx 250 × 350 nm) makes them ideal candidates for microscopic studies. Recombinant vaccinia viruses that express a viral envelope-specific, green fluorescent protein (GFP) chimera produce enveloped virions that fluoresce green. This fluorescent labeling allows the live, real-time study of viral egress using a variety of microscopic techniques. The methods presented here describe how to image the movement of intracellular enveloped virions that are labeled with green fluorescent protein using time-lapse laser scanning confocal microscopy. Details are also provided for analyzing the images obtained and converting them into QuickTime movies suitable for presentation.

Key Words

Vaccinia virus B5R green fluorescent protein (GFP) intracellular enveloped virus intracellular movement confocal microscopy time-lapse microscopy NIH image QuickTime