Plaquing of Herpes Simplex Viruses.

Lauren A Sadowski, Gregory M Lesko, Chad Suissa, Rista Upadhyay, Prashant J Desai, Barry J Margulies

Journal of visualized experiments : JoVE 2021 Nov 05

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There are numerous published protocols for plaquing viruses, including references within primary literature for methodology. However, plaquing viruses can be difficult to perform, requiring focus on its specifications and refinement. It is an incredibly challenging method for new students to master, mainly because it requires meticulous attention to the most minute details. This demonstration of plaquing herpes simplex viruses should help those who have struggled with visualizing the method, especially its nuances, over the years. While this manuscript is based on the same principles of standard plaquing methodology, it differs in that it contains a detailed description of (1) how best to handle host cells to avoid disruption during the process, (2) a more useful viscous medium than agarose to limit the diffusion of virions, and (3) a simple fixation and staining procedure that produces reliably reproducible results. Furthermore, the accompanying video helps demonstrate the finer distinctions in the process, which are frequently missed when instructing others on conducting plaque assays.

Citation

Lauren A Sadowski, Gregory M Lesko, Chad Suissa, Rista Upadhyay, Prashant J Desai, Barry J Margulies. Plaquing of Herpes Simplex Viruses. Journal of visualized experiments : JoVE. 2021 Nov 05(177)