TY - JOUR
T1 - Plaquing of herpes simplex viruses
AU - Sadowski, Lauren A.
AU - Lesko, Gregory M.
AU - Suissa, Chad
AU - Upadhyay, Rista
AU - Desai, Prashant J.
AU - Margulies, Barry J.
N1 - Funding Information:
We thank countless students in our labs (PJD and BJM) who have worked with us over the years refining these methods. A special thanks to Stan Person, under whose tutelage this methodology was first developed. This work was partially supported by the Towson University Fisher College of Science and Math Undergraduate Research Support fund and NIGMS Bridges to the Baccalaureate grant 5R25GM058264. This content is solely the authors' responsibility and does not necessarily represent the official views of the National Institutes of Health's National Institute of General Medical Sciences.
Publisher Copyright:
© 2021 JoVE Journal of Visualized Experiments.
PY - 2021/11
Y1 - 2021/11
N2 - 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.
AB - 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.
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U2 - 10.3791/61786
DO - 10.3791/61786
M3 - Article
C2 - 34842228
AN - SCOPUS:85121154169
SN - 1940-087X
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 177
M1 - e62962
ER -