title: Phytoplankton monitoring by high performance flow cytometry: a successful approach?
abstract: BACKGROUND
  Regular phytoplankton monitoring in Dutch coastal waters is performed as an indicator of the ecological state of these waters. The monitoring program is focused on temporal and spatial changes of species composition and abundance. Flow cytometry has been introduced to provide additional information, to improve ecosystem understanding, and to increase the efficiency of analysis and reportage.
  
  
  METHODS
  Phytoplankton community abundance and composition were routinely determined by flow cytometry and microscopy at six locations in the North Sea over three annual cycles between 2000 and 2003. Supplementary measurements were also made for fluorescence (chlorophyll-a and other pigments) and, in combination with flow cytometric and microscopic data, were used to determine phytoplankton abundance and composition as a function of their size distribution. Real-time imaging of species was also used to identify species on the basis of their flow cytometric optical characteristics.
  
  
  RESULTS
  Flow cytometric analysis took 15 min on average. Analysis including data processing, and Web site reportage took less than 1 h. Phytoplankton concentrations (cells/ml), biomass (fluorescence/ml), and concentration of phycoerythrin- or phycocyanin-containing cells (cells/ml) as a function of their algal size were produced every 2 weeks on average. The phytoplankton integrated annual concentration and biomass were used as ecological indicators for overall phytoplankton status. Real-time imaging of cells in flow enabled the identification of dominant species and was applied as an early warning system for Phaeocystis spp.
  
  
  CONCLUSIONS
  The reproducibility and count precision due to the large number of observations of the flow cytometric technique provided reliable data for monitoring long-term trends. Flow cytometrically based analyses extended the lower detection limit (<0.5 microm) of analysis beyond the capabilities of other techniques such as the relation between small and larger phytoplankton, the relation between cell counts and biomass as a function of cell size, but also the ability to monitor and report on blooms of harmful algae. A good correlation was found between concentrations (cells/ml) measured by flow cytometry and microscopy. In practice, flow cytometric analysis of a single marine sample took 15 min on average.
authors.affiliation: National Institute for Coastal and Marine Management, Middelburg, The Netherlands. t.p.a.rutten@rikz.rws.minvenw.nl
authors.forename: Thomas P A
authors.initials: TP
authors.lastname: Rutten
authors.forename: Ben
authors.initials: B
authors.lastname: Sandee
authors.forename: Angelo R T
authors.initials: AR
authors.lastname: Hofman
mesh_terms: D000818:Animals; D017753:Ecosystem; D004784:Environmental Monitoring / Q000191:economics / Q000379:methods; D005434:Flow Cytometry / Q000191:economics / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D008853:Microscopy / Q000191:economics / Q000379:methods; D009426:Netherlands; D010839:Phytoplankton; D011786:Quality Control