Dataset: BLOOFINZ-IO Flow Cytometry Abundance
Deployment: RR2201

Gating analyses of the listmode files followed those in Selph 2021 for the Hoechst-stained samples. For samples stained with Hoechst and LTG, the gating involved many steps as follows. LTG positive and negative particles were separated assuming that the maximum negative signal was that of co-occurring prokaryotes (Prochlorococcus, Synechococcus, and non-pigmented bacteria). Thus, all particles that had LTG signals greater than that of prokaryotes were designated LTG POS. LTG POS particles were further separated by the signal of the forward light scatter (FSC) of a 1 micrometer (µm) yellow-green fluorescent plastic beads (Polysciences, Inc.). Those particles with less FSC than a 1 µm bead were designated "PICO" sized particles and plotted as chlorophyll vs. phycoerythrin to eliminate any cyanobacteria, whereas the remaining cells were designated as potential mixotrophic cells. Those with more scatter than a 1 µm bead were designated "NANO+" sized particles and plotted as a single parameter chlorophyll histogram, where those with more chlorophyll than Prochlorococcus were considered potential mixotrophic eukaryotic cells. LTG POS PICO and NANO+ particles comprise the MEUK category reported. NANO+ (LTG POS) particles with less chlorophyll than Prochlorococcus were designated as heterotrophic eukaryotes (HEUK).