Enumerating algal viruses by flow cytometry v2 [dataset]

Amy Zimmerman, Camille Poirier
2016 protocols.io  
1 1 1 VERVE Net Worden Lab Purpose : Purpose : To quantify virus-like particles in culture or environmental samples. Summa ry: Summa ry: Host cells and virus-like particles are enumerated from the same glutaraldehyde-fixed sample. For virus counts, an aliquot is diluted in 0.02-µm-filtered TE buffer prior to staining with SYBR Green I nucleic acid dye to improve staining efficiency and ensure an appropriately low event rate in the instrument. An unstained aliquot of sample is run for host cell
more » ... ounts. Pri nci pl e Pri nci pl e Combining high-fluorescence-yield nucleic-acid-specific stains with flow cytometry has significantly improved the speed of analysis and accuracy of counting virus-like particles. Robust quantification using this method relies on reproducible staining efficiency as well as appropriate instrument parameters to ensure all events of interest are on scale. This protocol builds on the work of Brussaard (2004) Appl Environ Microbiol. (
doi:10.17504/protocols.io.guibwue fatcat:6qj2dtejrnfmrhdwyk2qxczwaq