Structure-Based Mutations in the Herpes Simplex Virus 1 Glycoprotein B Ectodomain Arm Impart a Slow-Entry Phenotype

Fan, Qing; Kopp, Sarah J.; Connolly, Sarah A.; Longnecker, Richard

Microscopy and determination of plaque size. Fluorescence microscopy of RFP expression in infected Vero cells was performed with an EVOS Cell Imaging System (AMG, Fisher Scientific). Plaques of both WT and gB 3A viruses were visualized by Giemsa (Sigma-Aldrich) staining after 3 days of incubation and then imaged by transmitted-light microscopy with an EVOS Cell Imaging System. Plaques size was calculated by using the EVOS Cell Imaging System. Randomly selected plaques were enlarged on a computer screen, and the average plaque radius was calculated from two independent measurements of at least 20 plaques. By using this average radius, the plaque area and the ratio of the plaque sizes of mutant and WT viruses were determined.