Methods Mol Biol. 2004;269:205-18.

Pox, dyes, and videotape: making movies of GFP-labeled vaccinia virus.

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Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.

Abstract

The large size of poxvirus virions (approx 250 x 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.

PMID:: 15114018; [PubMed - indexed for MEDLINE]

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