Suspended marine particles constitute most of the particulate organic matter pool in the oceans, thereby providing substantial substrates for heterotrophs, especially in the mesopelagic. Conversely, sinking particles are major contributors to carbon fluxes defining the strength of the biological carbon pump (BCP). This study is the first to investigate the differential influence of eukaryotic communities to suspended and sinking particles, using 18S rRNA gene sequencing on particles collected with a marine snow catcher in the mixed layer and upper mesopelagic of the Scotia Sea, Southern Ocean. In the upper mesopelagic, most eukaryotic phytoplankton sequences belonged to chain-forming diatoms in sinking particles and to prymnesiophytes in suspended particles. This suggests that diatom-enriched particles are more efficient in carbon transfer to the upper mesopelagic than those enriched in prymnesiophytes in the Scotia Sea, the latter more easily disintegrating into suspended particles. In the upper mesopelagic, copepods appeared most influential on sinking particles whereas soft-tissue metazoan sequences contributed more to suspended particles. Heterotrophic protists and fungi communities were distinct between mixed layer and upper mesopelagic, implying that few protists ride along sinking particles. Furthermore, differences between predatory flagellates and radiolarians between suspended and sinking particles implied different ecological conditions between the two particles pools, and roles in the BCP. Molecular analyses of sinking and suspended particles constitute powerful diagnostic tools to study the eukaryotic influence on the BCP in a more holistic manner compared to classic carbon export studies focusing on sinking particles.