title: Improvement of phytoplankton culture isolation using single cell sorting by flow cytometry.
abstract: Flow cytometry provides a tool to physically sort single algal cells in order to obtain clonal cultures. During sorting, cells are submitted to physical stress factors such as high fluidic pressure, exposure to the laser beam, electrostatic charges, deflection through high voltage fields, and collisions with container surfaces. All of these can damage the cells of interest and success rates for initiation of cultures from flow-sorted cells are generally very low. We found that the addition of bovine serum albumin in the culture medium into which cells were sorted drastically improved the success of initiation of pico- and nano-eukaryotic phytoplankton strains. Adding a mixture of antibiotics (Penicillin, Neomycin, Streptomycin) to the medium in order to slow down bacterial growth further improved culture development. This approach was successfully used to isolate taxonomically diverse strains, including novel taxa, from a fresh sample obtained in the English Channel and from enrichment cultures established during an Atlantic meridional transect cruise. We anticipate that these improvements will be useful to clone or purify existing cultures and to isolate novel cultures from oceanic samples.
authors.affiliation: UPMC Université Paris 06, CNRS, UMR7144, Station Biologique de Roscoff, Sorbonne Universités, Roscoff, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Marie
authors.affiliation: UPMC Université Paris 06, CNRS, UMR7144, Station Biologique de Roscoff, Sorbonne Universités, Roscoff, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Le Gall
authors.affiliation: UPMC Université Paris 06, CNRS, UMR7144, Station Biologique de Roscoff, Sorbonne Universités, Roscoff, France.
authors.forename: Roseline
authors.initials: R
authors.lastname: Edern
authors.affiliation: UPMC Université Paris 06, CNRS, UMR7144, Station Biologique de Roscoff, Sorbonne Universités, Roscoff, France.
authors.forename: Priscillia
authors.initials: P
authors.lastname: Gourvil
authors.affiliation: UPMC Université Paris 06, CNRS, UMR7144, Station Biologique de Roscoff, Sorbonne Universités, Roscoff, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Vaulot
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D005434:Flow Cytometry; D009355:Neomycin / Q000494:pharmacology; D010406:Penicillins / Q000494:pharmacology; D010839:Phytoplankton / Q000166:cytology* / Q000187:drug effects; D013307:Streptomycin / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D010406:Penicillins; D009355:Neomycin; D013307:Streptomycin
keywords: 
  BSA
  ; antibiotics; flow cytometry; microalgal cultures; nanoeukaryotes; phytoplankton; picoeukaryotes; single cell sorting