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.
Showing items related by title, author, creator and subject.
Coco, Giovanni; Murray, A. Brad; Green, Malcolm O.; Thieler, E. Robert; Hume, T. M. (American Geophysical Union, 2007-08-14)We employ a numerical model to study the development of sorted bed forms under a variety of hydrodynamic and sedimentary conditions. Results indicate that increased variability in wave height decreases the growth rate of ...
Greene, Charles H.; Meyer-Gutbrod, Erin; Monger, Bruce C.; McGarry, Louise P.; Pershing, Andrew J.; Belkin, Igor M.; Fratantoni, Paula S.; Mountain, David G.; Pickart, Robert S.; Proshutinsky, Andrey; Ji, Rubao; Bisagni, James J.; Hakkinen, Sirpa M. A.; Haidvogel, Dale B.; Wang, Jia; Head, Erica; Smith, Peter; Reid, Philip C.; Conversi, Alessandra (Association for the Sciences of Limnology and Oceanography, 2013-05)Decadal-scale regime shifts in Northwest Atlantic shelf ecosystems can be remotely forced by climate-associated atmosphere–ocean interactions in the North Atlantic and Arctic Ocean Basins. This remote climate forcing is ...
Green, Kenneth Edward (Massachusetts Institute of Technology and Woods Hole Oceanographic Institution, 1980-02)The measurements of the heat flow field of the Galapagos Spreading Center in an area of about 570 km2 reveal the planform of the conductive flux and permit the first truly areal estimate of the near-axis heat flux for ...