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Journal of Biomedical Optics • Open Access

Time-lapse ultrashort pulse microscopy of infection in three-dimensional versus two-dimensional culture environments reveals enhanced extra-chromosomal virus replication compartment formation
Author(s): Holly C. Gibbs; Garwin Sing; Juan Carlos Gonzalez Armas; Colin J. Campbell; Peter Ghazal; Alvin T. Yeh

Paper Abstract

The mechanisms that enable viruses to harness cellular machinery for their own survival are primarily studied in cell lines cultured in two-dimensional (2-D) environments. However, there are increasing reports of biological differences between cells cultured in 2-D versus three-dimensional (3-D) environments. Here we report differences in host-virus interactions based on differences in culture environment. Using ultrashort pulse microscopy (UPM), a form of two-photon microscopy that utilizes sub-10-fs pulses to efficiently excite fluorophores, we have shown that de novo development of extra-chromosomal virus replication compartments (VRCs) upon murine cytomegalovirus (mCMV) infection is markedly enhanced when host cells are cultured in 3-D collagen gels versus 2-D monolayers. In addition, time-lapse imaging revealed that mCMV-induced VRCs have the capacity to grow by coalescence. This work supports the future potential of 3-D culture as a useful bridge between traditional monolayer cultures and animal models to study host-virus interactions in a more physiologically relevant environment for the development of effective anti-viral therapeutics. These advances will require broader adoption of modalities, such as UPM, to image deep within scattering tissues.

Paper Details

Date Published: 1 March 2013
PDF: 7 pages
J. Biomed. Opt. 18(3) 031111 doi: 10.1117/1.JBO.18.3.031111
Published in: Journal of Biomedical Optics Volume 18, Issue 3
Show Author Affiliations
Holly C. Gibbs, Texas A&M Univ. (United States)
Garwin Sing, The Univ. of Edinburgh (United Kingdom)
Juan Carlos Gonzalez Armas, The Univ. of Edinburgh (United Kingdom)
Colin J. Campbell, The Univ. of Edinburgh (United Kingdom)
Peter Ghazal, The Univ. of Edinburgh (United Kingdom)
Alvin T. Yeh, Texas A&M Univ. (United States)

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