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These data represent the abundance, biovolume, and biomass of prokaryotic phytoplankton, eukaryotic pico- and nano- phytoplankton, and heterotrophic bacteria from discrete flow cytometry samples collected during the Northeast U.S. Shelf Long-Term Ecological Research (NES-LTER) Transect cruises, ongoing since 2018. Samples were collected and preserved from the water column at multiple depths using Niskin bottles on a CTD rosette system along the NES-LTER transect, and analyzed post cruise. Cells were identified and enumerated from the flow cytometry data files based on their scattering, SYBR (525 nm), phycoerythrin (575 nm) and chlorophyll (680 nm) fluorescence signals. Gating was completed manually in the Attune NXT software interface.