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JPR Advance Access published online on October 17, 2006
Journal of Plankton Research, doi:10.1093/plankt/fbl059
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© The Author 2006. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received June 8, 2006
Accepted October 12, 2006

Article

Flow cytometric enumeration of DNA-stained oceanic planktonic protists

Mikhail V. Zubkov 1 *, Peter H. Burkill 1, and Juliette N. Topping 1

1 National Oceanography Centre Southampton, European Way, Southampton, United Kingdom

* To whom correspondence should be addressed.
Mikhail V. Zubkov, E-mail: mvz{at}noc.soton.ac.uk


  Abstract

The aim of the present study was to test the practicality of enumerating fixed, DNA-stained heterotrophic protists (H) and phototrophic protists (P) in contrasting regions of the Atlantic Ocean. Oceanic protists were enumerated using a standard flow cytometer (FACSort, BD) at an enhanced flow rate of up to 1.0 mL min-1 to increase numbers of counted cells. The enumeration error of protists decreased hyperbolically from 30-40% to <5% corresponding to a number (<100 to >2000) of enumerated cells. H and P were discriminated using the extra red chlorophyll-derived plastidic fluorescence of the latter. The relationship between counts of stained and unstained fixed and unfixed P was statistically close to 1:1, confirming the accuracy of stained protist counting by flow cytometry and adequate discrimination of P from H cells. The estimated average abundance of H in the surface mixed layer of the Southern and Northern oligotrophic gyres was remarkably similar, with 400±140 and 450±60 cells mL-1, respectively, adding further evidence to the suggestion that these regions are in steady state. In agreement with earlier studies in more productive aquatic environments, a significant correlation (correlation coefficient 0.84, p<0.0001), was found between the H and total bacterioplankton numbers, with an average ratio ~1300 prokaryotes to 1 H cell, suggesting a relatively constant trophic interaction between these two groups. The present study demonstrates that flow cytometric enumeration of protists is ca 100 times faster compared to microscopy and, thus, represents a major improvement for quantifying protists in ocean waters, including oligotrophic gyres.

Communicating Editor: KJ Flynn


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