Earlier, we found with Epstein Barr virus and Type 2 Herpes simplex that copper acetate added after virus attachmment reduced significantly virion production as measured by immunofluorescence microscopy on fixed monolayers (Orsi et al 1988, Obradovic & Orsi 1990). However with similar conditions lead acetate did not modify virus replication. Our microscopy methods provided insufficient emission intensity for measuring the low levels of fluorescence associated with viral DNA formation and the early stages of virion maturation. We then devised a method based on procedures used for fixed cell visualization (Narin 1964, Brigati et al 1983). In our method, cell monolayers were infected, metal added, cells fixed and permeabilized for addition of the fluorescent probe which was then extracted and the emission intensity measured by fluorometry.
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- A Novel Fluorometric Method for Monitoring Virus Replication In Response to Heavy Metals
E. V. Orsi
- Springer Berlin Heidelberg