Fluorescent Cell-Counting Assay of Yellow Fever Virus

Cell Attachment and Penetration by Influenza Virus

“…For this procedure, virus dilutions were prepared in PBS (0.15 M NaCl, 0.01 M phosphate), pH 7.1. Inoculum in 0.2-ml volume was introduced directly onto cover-slip cell monolayers after their transfer from glass vials to rotor chamber inserts (10). The latter were employed because they withstand the centrifugal force required to sediment virus.…”

Assay of Chikungunya Virus in Cell Monolayers by Immunofluorescence

“…The outstanding feature of the method, in common with immunofluorescent assays described for other arboviruses, 2 , 5 wni the ability to assess virus infectivity within 24 hours after inoculation of cell monolayers.…”

“…1 Of the large number of agents that comprise the different arbovirus groups, however. immunofluorescent assays bave been developed for only four viruses: yellow fever, 2 Venezuelan equine encephalomyelitis, 3 Semliki Forest fever,6 and Rift Valley fever. 6 The feasibility of extending the immunofluorescent technique for the infectivity ass,:ssment of another arbovirus, chikungunya, was investigated because it has been demonstrated that cells infected with the virus are amenable to immunofluorescent staining.5"' This is a report on the development and evaluation of this technique for the quantitative assay of chikungunya virus.…”

Multiple Infection of Cell Monolayers by Virus Mixtures

“…Centrifugation (19,000 to 29,000 x g, 15 min) was employed for adsorption of inoculum onto cover slip cell cultures held in rotor chamber inserts. 7 Cell cultures were then incubated at 35 C for 16 to 24 hours in accordance with previously established assays for each agent.s' 9 After fixing with acetone at -60 C, cover slip cell monolayers were separated into three groups and selectively stained with one of three viral-immune sera conjugated with fluorescein isothiocyanate. The infected cells were then counted.…”