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    Comparison of techniques used to count single-celled viable phytoplankton

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    Phytoplankton_Workshop_JAppPhyco_revised_with-figures&tables.pdf (441.8Kb)
    Date
    2010-10-14
    Author
    Steinberg, Mia K.  Concept link
    First, Matthew R.  Concept link
    Lemieux, Edward J.  Concept link
    Drake, Lisa A.  Concept link
    Nelson, Bruce N.  Concept link
    Kulis, David M.  Concept link
    Anderson, Donald M.  Concept link
    Welschmeyer, Nicholas A.  Concept link
    Herring, Penny R.  Concept link
    Metadata
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    Citable URI
    https://hdl.handle.net/1912/5388
    As published
    https://doi.org/10.1007/s10811-011-9694-z
    Keyword
     Phytoplankton; Enumeration; FlowCAM; Flow cytometry; Sedgewick Rafter; Ballast water; SYTOX Green; CellTracker Green 
    Abstract
    Four methods commonly used to count phytoplankton were evaluated based upon the precision of concentration estimates: Sedgewick Rafter and membrane filter direct counts, flow cytometry, and flow-based imaging cytometry (FlowCAM). Counting methods were all able to estimate the cell concentrations, categorize cells into size classes, and determine cell viability using fluorescent probes. These criteria are essential to determine whether discharged ballast water complies with international standards that limit the concentration of viable planktonic organisms based on size class. Samples containing unknown concentrations of live and UV-inactivated phytoflagellates (Tetraselmis impellucida) were formulated to have low concentrations (<100 ml-1) of viable phytoplankton. All count methods used chlorophyll a fluorescence to detect cells and SYTOX fluorescence to detect non-viable cells. With the exception of one sample, the methods generated live and non-viable cell counts that were significantly different from each other, although estimates were generally within 100% of the ensemble mean of all subsamples from all methods. Overall, percent coefficient of variation (CV) among sample replicates was lowest in membrane filtration sample replicates, and CVs for all four counting methods were usually lower than 30% (although instances of ~60% were observed). Since all four methods were generally appropriate for monitoring discharged ballast water, ancillary considerations (e.g., ease of analysis, sample processing rate, sample size, etc.) become critical factors for choosing the optimal phytoplankton counting method.
    Description
    Author Posting. © The Author(s), 2010. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Journal of Applied Phycology 24 (2012): 751-758, doi:10.1007/s10811-011-9694-z.
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    • Biology
    Suggested Citation
    Preprint: Steinberg, Mia K., First, Matthew R., Lemieux, Edward J., Drake, Lisa A., Nelson, Bruce N., Kulis, David M., Anderson, Donald M., Welschmeyer, Nicholas A., Herring, Penny R., "Comparison of techniques used to count single-celled viable phytoplankton", 2010-10-14, https://doi.org/10.1007/s10811-011-9694-z, https://hdl.handle.net/1912/5388
     

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