Carbon biomass estimates of eukaryotic phytoplankton by taxa and size-class, based on epifluorescence microscopy from samples collected on R/V Melville cruise MV1008 in the Costa Rica Dome in 2010 (CRD FLUZiE project) (NCEI Accession 0278242)

Acquisition Description:
Seawater samples (500 mL) for microscopical analysis were gently collected from the CTD and immediately preserved for slide preparation according to a modified protocol of Sherr and Sherr (1993). The samples were first preserved with 260 uL of alkaline Lugol’s solution, immediately followed by 10 mL of buffered formalin and 500 uL of sodium thiosulfate, with gentle mixing between each addition. Preserved samples were shielded from light and left to rest at room temperature for 1 h. After the rest period, 1 mL of proflavin (0.33% w/v) was added and the samples were stored in the dark for an additional hour. Just prior to filtration, the preserved samples were stained with 1 mL of DAPI (0.01 mg mL -1 ) and immediately transferred to the filtration manifold. A 50-mL aliquot (small volume, SV) of the sample was filtered through a 25-mm black polycarbonate filter with 0.8-um pore size, and the remaining 450 mL aliquot (large volume, LV) was filtered through a 25-mm black polycarbonate filter with 8.0-um pores. A 10-mm nylon backing filter was placed under all polycarbonate filters to promote even cell distribution, and filtered the samples under gentle vacuum pressure (<100 mm Hg). Each filter was then mounted onto glass slides with one drop of Type DF immersion oil and a No. 2 cover slip, and the prepared slides were frozen at -80°C for later analysis in the lab.