Short communication
Sub-viral imaging of vaccinia virus using super-resolution microscopy

https://doi.org/10.1016/j.jviromet.2012.07.003Get rights and content

Abstract

The study of host–pathogen interactions over past decades has benefited from advances in microscopy and fluorescent imaging techniques. A particularly powerful model in this field is vaccinia virus (VACV), which due to its amenability to genetic manipulation has been a productive model in advancing the understanding of the transport of subcellular cargoes. Conventional light microscopy imposes an upper limit of resolution of ∼250 nm, hence knowledge of events occurring at the sub-viral resolution is based predominantly on studies utilising electron microscopy. The development of super-resolution light microscopy presents the opportunity to bridge the gap between these two technologies. This report describes the analysis of VACV replication using fluorescent recombinant viruses, achieving sub-viral resolution with three-dimensional structured illumination microscopy. This is the first report of successfully resolving poxvirus particle morphologies at the scale of single virus particles using light microscopy.

Highlights

► Imaging of single vaccinia virus particles at sub-viral resolution was achieved using 3D-SIM. ► The viral envelope protein B5 was resolved as a spherical structure encompassing the virus cores. ► Maturation and morphogenesis of virus particles is now accessible to light microscopy analysis.

Section snippets

Acknowledgements

This work was supported by the Australian Research Council Federation Discovery Project (DP1096623). CBW is supported by an Australian National Health and Medical Research Council Senior Research Fellowship. We would like to thank the members of the Newsome Lab for their helpful comments.

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