Abstract
Flow sorting can be a very helpful tool in revealing phytoplankton and bacterial community structure and elaborating specific physiological parameters of isolated species. Droplet sorting has been the most common technique. Despite the high optical and hydro-dynamic stress for the cells to be sorted, many species grow in culture subsequent to sorting. To date, flow sorting has been applied to post-incubation separation in natural water samples to account for group-specific physiological parameters (radiotracer-uptake rates), to the production of clonal or non-clonal cultures from mixtures, to the isolaton of cell groups from natural assemblages for molecular analyses, and for taxonomic identification of sorted cells by microscopy. The application of cell sorting from natural water samples from the Wadden Sea, including different cryptophytes, cyanobacteria and diatoms, is shown, as well as the establishment of laboratory cultures from field samples. The optional use of a red laser to account for phycocyanine-rich cells is also discussed.
Highlights
The essence of flow cytometry is the simultaneous measurement of different optical cell properties, which allow the characterization and classification of individual cells in a mixture (Hofstraat et al, 1991; Veldhuis and Kraay, 2000)
The autofluorescence of the naturally occurring chlorophylls and phycobilins in phytoplankton set the natural premise for the application of flow cytometry, and it was not long before flow cytometry became a widely used tool in characterizing marine phytoplankton assemblages (Yentsch and Horan, 1989)
The cryptophyte genera Hemiselmis is interesting in that it is the only cryptophyte genus with some species possessing phycoerythrine and others phycocyanine (Hill and Rowan, 1989). In addition to these clusters, which are characterized by their specific phycobilin fluorescence, other eukaryotic phytoplankton were differentiated by their light scatter and red fluorescence properties
Summary
SUMMARY: Flow sorting can be a very helpful tool in revealing phytoplankton and bacterial community structure and elaborating specific physiological parameters of isolated species. Despite the high optical and hydro-dynamic stress for the cells to be sorted, many species grow in culture subsequent to sorting. Flow sorting has been applied to post-incubation separation in natural water samples to account for group-specific physiological parameters (radiotracer-uptake rates), to the production of clonal or non-clonal cultures from mixtures, to the isolaton of cell groups from natural assemblages for molecular analyses, and for taxonomic identification of sorted cells by microscopy. The application of cell sorting from natural water samples from the Wadden Sea, including different cryptophytes, cyanobacteria and diatoms, is shown, as well as the establishment of laboratory cultures from field samples. The optional use of a red laser to account for phycocyanine-rich cells is discussed
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