Techniques for quantitative analyses of calcareous marine phytoplankton

This paper discusses the techniques used to sample and analyse living marine calcareous phytoplankton. The various methods are described and tested within several research projects aimed at the determination of coccolithophore cell densities in seawater. In addition, the potential advantages and drawbacks associated with the application of light and scanning electron microscopic techniques to the quantitative analysis of coccolithophores are discussed. Several tests have been carried out in order to quantify potential errors related to: (1) homogeneity of material distribution on filter membranes; (2) use of different microscopes (scanning electron microscope versus light microscope); (3) use of different filter membranes (cellulose mixed-ester membranes versus polycarbonate membranes); and (4) Utermöhl settling versus filtration method. These tests revealed that major errors in cell density calculations could result from the uneven distribution of coccolithophore specimens on a filter membrane. The error resulting from the use of a light microscope arises from its low resolution, which restricts the identification of species, especially of small coccospheres. The use of different filter membranes does not show a statistically significant difference in cell density calculations, although polycarbonate membranes can be examined much more efficiently with the scanning electron microscopy than cellulose mixed-ester membranes. The Utermöhl method, however, gives lower cell densities consistently (several times) than the filtration method.