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Application of Imaging Flow Cytometry to Monitor Life Cycle Transitions in Alexandrium tamarense species Shahla Farzan1, Michael L. Brosnahan2, Mireia Lara Artigas3, Robert J. Olson2, and Donald M. Anderson2 1. Mount Holyoke College, South Hadley, MA, 01075, U.S.A. 2. Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA, 02543, U.S.A. 3. Instituto Oceanogràfico Vigo, Cabo Estai-Canido, 36200 Vigo, Spain Alexandrium tamarense are globally distributed dinoflagellate species that may produce toxins associated with paralytic shellfish poisoning (PSP). Previous research has shown that two A. tamarense species (Group I and Group III) can interbreed in culture and in nature, but the resulting progeny of these hybrids are inviable. These findings are particularly interesting because Group I cells produce PSP toxins while Group III cells do not. Therefore, the introduction of nontoxic cells into the marine environment could be an effective means of mitigating toxic A. tamarense blooms. Before such a strategy can be pursued, a more complete understanding of the factors governing sexuality is needed. In general, light microscopy is used to monitor life cycle transitions. However, this method is inefficient and possibly unreliable because sexual stages are difficult to differentiate from asexual vegetative cells. To address the problem of low sample throughput, we have configured the Imaging Flow Cytobot (IFCB), a submersible flow cytometry system, for automated detection of A. tamarense cells that have been stained with a Group I specific ribosomal probe. The system measures cellular DNA content in order to confirm diploidy in swimming planozygote cells. The IFCB system will be used to assess the appearance and overall abundance of sexual stages in a series of mating experiments between Group I and Group III clones. A modified IFCB will also be used to analyze samples taken over the course of a natural bloom in the Nauset Marsh area (Cape Cod, MA).
