Immuno-flow cytometric identification and enumeration of the ichthyotoxic dinoflagellate Gyrodinium aureolum Hulburt in artificially mixed algal populations

Flow cytometric identification and enumeration of Gyrodiniumaureolum Hulburt (Dinophyceae) were performed in artiticiallymixed algal populations using direct immunofluorescence. Calibrationof the flow cytometer, performed with a mixed algal populationspiked with immuno-fluorescently labelled G.aureolum cells,showed that selection of target cells after analysis on greenand orange fluorescence can be done with a recovery of 91.8%[coefficient of variation (CV) = 0.09]. Other selection methodswere less good, with 67.4% (CV = 0.16) and 58.4% (CV = 0.43)recovery based on green and red fluorescence or green fluorescenceand perpendicular light scattering. For mixed algal populationsspiked with unlabelled G.aureolum cells, the quantificationof target cells was quite good (recovery of 76.7%; CV = 0.20).The percentage of total cell loss was high (58.0–92.0%),but this was caused mostly by loss of species smaller in sizethan G.aureolum. Estimates of the relative contribution of G.aureolumin labelled samples were therefore often far too high, but detectionand quantification were not affected. The methodological underestimation(23.3%) was partly caused by gating on green and orange fluorescence(inaccuracy 8.2%).