Quantification of Viable but Non-Culturable Cells of Legionella pneumophila
Legionella pneumophila, among other bacteria, may enter a viable but non-culturable state as a means for survival in stressful conditions. Bacterial cells in the viable but non-culturable state cannot grow on standard medium; however, they continue to exhibit characteristics that are associated with live cells, such as respiration, transcription, and cell wall integrity. The present paper outlines a detailed protocol for the detection of viable but non-culturable L. pneumophila cells via Syto® 9 and propidium iodide staining coupled with flow cytometry.
Key wordsViable but non-culturable state Staining and labeling Flow cytometry Legionella pneumophila
This work was supported by Discovery Grant 418289-2012 from the National Sciences and Engineering Research Council of Canada (NSERC) and a John R. Evans Leaders Fund—Funding for research infrastructure from the Canadian Foundation for Innovation to SPF. NM was the recipient of a PhD scholarship from Fond de Recherche du Québec—Nature et Technologie.
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