Abstract
Flow cytometry has been instrumental in characterizing normal and infected cells. However, until recently, it was not possible to use such an approach to analyze small entities such as bacteria, let alone viruses, owing to the 0.5 μm resolution of most instruments. To circumvent this limitation, some laboratories decorate pathogens with antibodies or nanoparticles. Our laboratory instead exploits an alternative approach that relies on the staining of internal viral constituents with permeable SYTO dyes or the fluorescent tagging of individual viral proteinaceous components, whether capsid, tegument or glycoproteins. This opens up a range of new research avenues and, for example, enabled us to characterize individual herpes simplex virus type 1 particles, discern their different subpopulations, measure the heterogeneity of mature virions in terms of protein content, sort these viral particles with >90% purity and, for the first time, directly address the impact of this heterogeneity on viral fitness. This approach, coined flow virometry or nanoscale flow cytometry, allows for the study of a wide variety of pathogens with high statistical significance and the potential discovery of novel virulence factors.
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Acknowledgments
This work was supported by grants from the Canadian Institutes of Health Research (MOP 82921). Special thanks to Diane Gingras for the optimization of the EM protocol and Annie Gosselin for her help with the flow cytometry section. We are also indebted to Daniele Gagné, who initially worked out the protocol on the BD FACSAria sorter.
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Khadivjam, B., El Bilali, N., Lippé, R. (2020). Analysis and Sorting of Individual HSV-1 Particles by Flow Virometry. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus . Methods in Molecular Biology, vol 2060. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9814-2_16
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DOI: https://doi.org/10.1007/978-1-4939-9814-2_16
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