Zooplankton samples were collected during the RV Meteor cruise M174 (April–May 2021) using a Multi Plankton Sampler (Multinet Midi, 200 µm mesh size). Sampling included oblique tows at discrete depth intervals (0–25, 25–50, 50–100, and 100–200 m), integrated oblique tows (0–200 m), and horizontal tows at selected stations. The cruise surveyed coastal and oceanic waters under and beyond the influence of the Amazon River Plume. Zooplankton samples were separated into two size fractions (>1 mm and <1 mm), digitized using a ZooScan imaging system (2400 dpi resolution), and processed with ZooProcess software. Images were subjected to automated classification followed by manual taxonomic validation using the EcoTaxa web application. Data were subjected to quality control in accordance with the protocols of the Pelagic Size Structure Database (PSSdb; https://pssdb.net/). Morphometric measurements obtained from scanned images were used to estimate individual carbon biomass (mg C) from image-derived organism projected area using published taxon-specific size–biomass and dry weight-to-carbon conversion relationships. Biomass estimates were subsequently standardized by the sampled water volume. When multiple tows sampled the same depth interval, biomass values were combined using midpoint integration to obtain weighted mean estimates representative of the corresponding depth interval. This dataset provides taxon-specific mesozooplankton biomass (mgC/m³) for samples collected along a gradient of Amazon River Plume influence across multiple depth strata during cruise M174.