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The copepod Pseudodiaptomus marinus<\/em> was cultured at RTC and used in the experiments. The evening before an experiment the copepods were size fractioned to separate the adults from the nauplii (larval stage) and were placed in 0.45 \u00b5m filtered San Francisco Bay water with the alga Rhodomonas salina, <\/em>to allow for feeding.<\/em> We handpicked 25 adults and 50 nauplii (Naupliar stage 3 and 4) which were maintained in 3 L of previously described water. The following morning (0900) the copepods were rinsed into freshly filtered 0.45 \u00b5m San Francisco Bay water and held for three hours to allow the individuals to clear the contents of their guts. While the copepods cleared their guts, the ciliates (Codonellopsis sp. <\/em>or Favella sp.<\/em>) were stained with the fluorescent stain, CellTracker\u2122 Green CMFDA (5-chloromethylfluorescein diacetate) (Li et al. 1996) for one hour at 1 \u00b5molar solution. After staining was complete, the ciliates were rinsed from the stain by being rinsed over a 35\u00b5m mesh and gently dunked into clean 0.45 \u00b5m filtered San Francisco Bay water. The ciliates were then resuspended and introduced to the copepods, which were allowed to feed for 1 hour.<\/p>\n To obtain the best images of the copepod digestive track the individuals needed to be alive. After 1 hour of feeding the copepods were rinsed over a 100 \u00b5m mesh which allowed for the separation of the ciliates and copepods. Using a dissecting microscope, individuals were transferred to a microscope slide in a small amount of water, which held each individual on its side for the best image of the gut. Once in place the microscope slide was transferred to the Olympus IX83 Epifluorescence microscope, and each individual was imaged under two settings. The first setting was bright-field and the second setting was the GFP filter (395 nm excitation, 500 nm emission) which allowed for the detection of the stained ciliate in the digestive track. This experiment was repeated twice.<\/p>\n Our next approach to identify the grazing of copepods on ciliates focused on offering the copepods a natural assemblage of food that was spiked with a known amount of stained ciliates. Three liters of surface seawater was collected from the seawall at RTC, size fractioned by reverse filtration into a 20-50 \u00b5m cohort, which removed other large grazers yet maintained other species of ciliates and phytoplankton. Cultured Favella sp.<\/em> was stained as previously described and then introduced to the natural assemblage. Two containers (1500 ml each) were made and the copepods were allowed to feed for 1 hour. At the end of the grazing period 3 replicate samples of 50 ml each were preserved with 0.1% Acid Lugol and 3 replicate samples of 50 ml each were preserved in 50% Glutaraldehyde.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/ocean-data.org\/schema\/hasBriefDescription":[{"@value":"Pseudodiaptomus marinus gut minimum fluorescence thresholds","@language":"en-US"}],"http:\/\/purl.org\/dc\/terms\/description":[{"@value":" Minimum fluorescence thresholds were measured for adult and nauplius\u00a0Pseudodiaptomus marinus<\/em> guts.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/www.w3.org\/2000\/01\/rdf-schema#label":[{"@value":"4b_min fluorescence thresholds","@type":"xsd:string"}],"http:\/\/ocean-data.org\/schema\/hasProcessingDescription":[{"@value":" The images were analyzed using Photoshop and Image J to calculate a Gut Fullness Index which was calculated as a percent. All images were processed identically including, calculation of total gut volume, a threshold correction to account for autofluorescence, and amount of gut full of the food item.\u00a0<\/p>\n BCO-DMO Processing:<\/strong>
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