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Triplicate 5 mL samples were preserved for flow cytometry with 0.5% glutaraldehyde (final concentration), incubated at 4\u00b0C for 10 min and frozen (-80\u00b0C) until analysis (within 2-3 weeks; Kemp et al. 1993). To calculate phytoplankton group abundances, 200 \u00b5l aliquots of fixed sample were added to a 96-well plate and run on a Guava flow cytometer (Millipore). Filtered seawater (0.45 \u00b5m) was run as a blank\u00a0and instrument-specific beads were used to calibrate the cytometer. Samples were analyzed at low flow rate (0.24 \u00b5l s-1) for 3 min. Three major phytoplankton groups were distinguishable based on plots of forward scatter vs. orange (phycoerythrin-containing, Synechococcus spp.) or red (pico- and nanoeukaryotes) fluorescence signals (Worden and Binder 2003).\u00a0<\/p>\n
Samples for enumerating bacteria were stained prior to running on the Guava in 0.5% v\/v SybrGreen I DNA stain for 1 hour at room temperature in the dark.<\/p>\n
Mesocosm treatment for all HHQ experiments was as follows:
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HHQ treatments here are as follows:
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\nDMSO control - equivalent (v:v) DMSO added to triplicate 5L bottles.<\/p>\n
\u00a0All bottles were incubated for 24h in a flow-through tank, that was shaded to mimic in situ conditions. Chlorophyll samples were taken at T0 and T24 for all experiments.<\/p>\n
Data were processed in Excel with statistics run in Excel, R, or Matlab.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/ocean-data.org\/schema\/hasBriefDescription":[{"@value":"HHQ Flow cytometry","@language":"en-US"}],"http:\/\/purl.org\/dc\/terms\/description":[{"@value":"
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BCO-DMO Processing Notes:<\/strong>
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