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

Abstract

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Physiological and behavioral diagnostics of nitrogen limitation for the toxic dinoflagellate Alexandrium fundyense

“…Our studies demonstrated that the use of chlorophyll fluorescence in conjunction with an antibody probe allows a population to be identified and sorted, but accurate enumeration was not achieved. As reported by Vrieling et al (1995) for Gyrodinium aureolum, we encountered significant cell loss during processing of samples for flow cytometric analysis. We determined that there are direct cell losses associated with the incubations and wash steps, but also found that inaccurate cell counts occurred because larger cells (e.g., A. fundyense) would partially block the flow cytometer sample tubing.…”

“…Vrieling et al 1994 andVrieling et al 1995). In this study we provide an evaluation of two antibody-based approaches to the isolation, enumeration, separation and biochemical analysis of a target species -the toxic dinoflagellate Alexandrium fundyense -in natural plankton samples.…”

“…(Costas and Lopez-Rodas 1996;Sako et al 1996), Gyrodinium aureolum and Gymnodinum sp. (Vrieling et al 1994;Vrieling et al 1995;Vrieling and Anderson 1996;Vrieling et al 1996;Vrieling et al 1997 (Table 2.1). Since more steps are involved in the labeling process with biotin-avidin with no significant signal enhancement, it is not worth the extra effort to use this labeling approach, at least for A.…”

“…However, larger size classes of phytoplankton, which include many toxic or harmful dinoflagellate species, often cannot be discriminated from other phytoplankton species based on size and autofluorescence alone. Therefore, molecular probes, specifically antibodies, are now used to tag or highlight toxic species for better discrimination (Hofstraat et al 1994;Vrieling et al 1994;Vrieling et al 1995). Some flow cytometers have cell sorting devices that allow physical separation of selected cells (gated cell populations) from the main sample stream.…”

“…However, larger size classes of phytoplankton which include toxic dinoflagellate species, cannot be discriminated as-easily based on size and autofluorescence alone. Therefore, antibody labeling procedures are now used to identify toxic phytoplankton species in the field (Vrieling et al 1994;Vrieling et al 1995;Vrieling et al 1995a;Poulton, unpub. data).…”