Comparison of techniques used to count single-celled viable phytoplankton
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.
MetadataShow full item record
KeywordPhytoplankton; Enumeration; FlowCAM; Flow cytometry; Sedgewick Rafter; Ballast water; SYTOX Green; CellTracker Green
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.
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.
Suggested CitationPreprint: 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
Showing items related by title, author, creator and subject.
Insights into the loss factors of phytoplankton blooms : the role of cell mortality in the decline of two inshore Alexandrium blooms Choi, Chang Jae; Brosnahan, Michael L.; Sehein, Taylor R.; Anderson, Donald M.; Erdner, Deana L. (2017-01)While considerable effort has been devoted to understanding the factors regulating the development of phytoplankton blooms, the mechanisms leading to bloom decline and termination have received less attention. Grazing ...
Lupton, John E.; Pyle, Douglas G.; Jenkins, William J.; Greene, Ronald; Evans, Leigh (American Geophysical Union, 2004-01-17)Several hydrographic stations in the vicinity of the Samoa Islands have 3He/4He above the regional background in the depth range of 1500–1800 m, indicating injection of mantle helium from a local hydrothermal source. The ...
Biomarkers signal contaminant effects on the organs of English sole (Parophrys vetulus) from Puget Sound Malins, Donald C.; Anderson, Katie M.; Stegeman, John J.; Jaruga, Pawel; Green, Virginia M.; Gilman, Naomi K.; Dizdaroglu, Miral (National Institute of Environmental Health Sciences, 2006-06)Fish living in contaminated environments accumulate toxic chemicals in their tissues. Biomarkers are needed to identify the resulting health effects, particularly focusing on early changes at a subcellular level. We used ...