Plaque forming unit assays were used to quantify the release of infectious viral particles [23 (link),28 (link)]. Vero cells were seeded the previous day in 48-well culture plates at a density of 3×104 cells per well. Cells were infected by 0.1 mL of ten-fold dilutions of supernatant. Following an incubation of 2 h at 37 °C, 0.2 mL of the culture medium supplemented with 5% fetal bovine serum (FBS) and 0.8% carboxymethylcellulose sodium salt (Sigma-Aldrich, Saint-Quentin-Fallavier, France)) were added, and the incubation was extended for 4 days at 37 °C. Cells were fixed (PFA, 3.7%) and stained with 0.5% crystal violet (Sigma-Aldrich) diluted in 20% ethanol, after the media had been removed. Plaques were counted and expressed as plaque-forming unit per mL (PFU mL−1).
Clain E., Haddad J.G., Koishi A.C., Sinigaglia L., Rachidi W., Desprès P., N. Duarte dos Santos C., Guiraud P., Jouvenet N, & El Kalamouni C. (2019). The Polyphenol-Rich Extract from Psiloxylon mauritianum, an Endemic Medicinal Plant from Reunion Island, Inhibits the Early Stages of Dengue and Zika Virus Infection. International Journal of Molecular Sciences, 20(8), 1860.
Other organizations :
Processus Infectieux en Milieu Insulaire Tropical, Institut de Recherche pour le Développement, Centre National de la Recherche Scientifique, Inserm, Fundação Oswaldo Cruz, Fundação Carlos Chagas, Sorbonne Paris Cité, Université Paris Cité, Institut Pasteur, Délégation Paris 7, CEA Grenoble, Institut Nanosciences et Cryogénie, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Université Grenoble Alpes
Culture medium (supplemented with 5% FBS and 0.8% carboxymethylcellulose sodium salt)
Incubation time after adding culture medium (4 days)
Incubation temperature after adding culture medium (37°C)
Fixation (PFA, 3.7%)
Staining (0.5% crystal violet in 20% ethanol)
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