<div><p><strong>Analytical procedure:</strong></p>
<p>Microbial cell abundances were determined using an Influx flow cytometer. Briefly, pigmented groups (Prochlorococcus, Synechococcus, and picoalgae) were enumerated in unstained samples by their chlorophyll and forward scatter signatures (Marie et al. 1999). The high phycoerythrin signal in Synechococcus was used to distinguish this group from Prochlorococcus and picoalgae.</p>
<p>To visualize non-pigmented picoplankton (bacteria) and heterotrophic protists, samples were stained with SYBR Green I DNA dye (SG, 1:10000 final concentration, for 10 min at 4C in the dark) (Zubkov et al. 2007, Christaki et al. 2011). Because bacteria and Prochlorococcus groups exhibit overlapping signal characteristics in surface samples after SG staining, the abundances of bacteria in surface samples were calculated by subtracting the Prochlorococcus abundance, determined in the unstained aliquot, from the total SG-stained group abundance. An internal standard of 1-um diameter fluorescent microspheres (Fluoresbrite, Polysciences) was added to each sample.</p>
<p><strong>Sampling and Analytical Methodology: </strong></p>
<p>Seawater samples were acquired from bottle samples collected by a CTD rosette during the OUTPACE cruise. 1.8 ml water samples were collected in duplicate, fixed with 0.25% (w/v) paraformaldehyde, flash frozen, and preserved at -80C. One of the duplicate samples was used to count phytoplankton (Prochlorococcus, Synechococcus, picoalgae), and the other was used to count bacteria and protists.</p>
<p>For pigmented groups (Prochlorococcus, Synechococcus, and picoalgae), cells were excited using a combination of two lasers (488+456 nm) focused into one pinhole; while for non-pigmented groups (bacteria and heterotrophic protists), cells were excited using the 488 nm laser only. For calibration, a solution of 1-um diameter fluorescent microspheres (Fluoresbrite, Polysciences) was used.</p>
<p><strong>References: </strong></p>
<p>Christaki U, Courties C, Massana R, Catala P, Lebaron P, Gasol JM, Zubkov MV (2011) Optimized routine flow cytometric enumeration of heterotrophic flagellates using SYBR Green I. Limnology and Oceanography-Methods 9:329-339. <a href="https://doi.org/10.4319/lom.2011.9.329">https://doi.org/10.4319/lom.2011.9.329</a></p>
<p>Marie D, Partensky F, Vaulot D, Brussaard C (1999) Enumeration of Phytoplankton, Bacteria, and Viruses in Marine Samples. Current Protocols in Cytometry:11.11.11-11.11.15. <a href="https://doi.org/10.1002/0471142956.cy1111s10">https://doi.org/10.1002/0471142956.cy1111s10</a></p>
<p>Zubkov M, Burkill PH, Topping JN (2007) Flow cytometric enumeration of DNA-stained oceanic planktonic protists. Journal of Plankton Research 29:79-86. <a href="https://doi.org/10.1093/plankt/fbl059">https://doi.org/10.1093/plankt/fbl059</a></p></div>
Flow cytometry cell counts obtained during the OUTPACE cruise
<div><p>This dataset contains abundances of cyanobacteria (<em>Synechococcus</em>, and <em>Prochlorococcus</em>), picoalgae, non-pigmented picoplankton (bacteria), and heterotrophic protists acquired through flow cytometry. Latitude, longitude, timestamp, and sample depth are also included in this dataset. </p>
<p>Water samples were acquired during the Oligotrophy to UlTra-oligotrophy PACific Experiment (OUTPACE) cruise between New Caledonia and Tahiti from February to April 2015.</p></div>
OUTPACE - flow cytometry cell counts
<div><p>Flow cytometry data were analyzed using FCS Express Pro (De Novo Software). </p>
<p>BCO-DMO Processing Notes:</p>
<p>* added ISO timestamp from OUTPACE cruise CTD logs <br />
* stripped out_c_ from station column for matching with bottle data <br />
* column station renamed cast to be consistent with CTD log<br />
* added a conventional header with dataset name, PI name, version date<br />
* modified parameter names to conform with BCO-DMO naming conventions</p></div>
663918
OUTPACE - flow cytometry cell counts
2016-11-03T16:47:32-04:00
2016-11-03T16:47:32-04:00
2023-07-07T16:10:26-04:00
urn:bcodmo:dataset:663918
Flow cytometry cell counts obtained during the R/V L'Atalante OUTPACE cruise between New Caledonia and Tahiti from February to April 2015
false
Duhamel, S. (2016) Flow cytometry cell counts obtained during the R/V L'Atalante OUTPACE cruise between New Caledonia and Tahiti from February to April 2015. Biological and Chemical Oceanography Data Management Office (BCO-DMO). Version Date 2016-11-03 [if applicable, indicate subset used]. http://lod.bco-dmo.org/id/dataset/663918 [access date]
true
false
2016-11-03
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