title: Rapid flow cytometry--Nile red assessment of PHA cellular content and heterogeneity in cultures of Pseudomonas aeruginosa 47T2 (NCIB 40044) grown in waste frying oil.
abstract: The accumulation of cytoplasmic polyhydroxyalkanoates (PHAs) and the heterogeneity ofbacterial populations were analysed by flow cytometry and SYTO-13 and Nile red staining in rhamnolipid-producing Pseudomonas aeruginosa cultures grown in waste frying oil as carbon source. A combination of SYTO-13 and Nile red fluorescence with cytometric forward and side scatter values may allow increases in the final production of polyhydroxyalkanoates (PHA) by two basic mechanisms: (i) rapid assessment of polyhydroxyalkanoate content and (ii) definition of flow cytometric cell sorting protocols to select high polyhydroxyalkanoate (PHA)-producing strains. We report a rapid (less than 30 min) flow cytometric assessment of PHAs in Pseudomonas aeruginosa 47T2 following Nile red staining: (i) to estimate cellular PHAs content; (ii) to study heterogeneity of the batch cultures producing PHAs and (iii) to establish the basis for sorting sub-populations with a high capacity to accumulate PHAs.
authors.affiliation: Department de Microbiologia i Parasitologia Sanitàries, Facultat de Farmacia, Universitat de Barcelona, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Vidal-Mas
authors.lastname: Resina-Pelfort
authors.forename: E
authors.initials: E
authors.lastname: Haba
authors.forename: J
authors.initials: J
authors.lastname: Comas
authors.forename: A
authors.initials: A
authors.lastname: Manresa
authors.forename: J
authors.initials: J
authors.lastname: Vives-Rego
mesh_terms: D003296:Cooking; D003470:Culture Media; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism*; D006017:Glycolipids / Q000378:metabolism; D008854:Microscopy, Electron; D010078:Oxazines / Q000378:metabolism*; D010938:Plant Oils; D011091:Polyesters / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000254:growth & development* / Q000378:metabolism; D013997:Time Factors
chemical_list: D003470:Culture Media; D005456:Fluorescent Dyes; D006017:Glycolipids; D010078:Oxazines; D010938:Plant Oils; D011091:Polyesters; C418382:rhamnolipid; C044808:nile red