In the Southern Ocean (SO), iron (Fe) limitation strongly inhibits phytoplankton growth and generally decreases their primary productivity. Diatoms are a key component in the carbon (C) cycle, by taking up large amounts of anthropogenic CO2 through the biological carbon pump. In this study, we investigated the effects of Fe availability (no Fe and 4 nM FeCl3 addition) on the physiology of Chaetoceros cf. simplex, an ecologically relevant SO diatom. Our results are the first combining oxygen evolution and uptake rates with particulate organic carbon (POC) build up, pigments, photophysiological parameters and intracellular trace metal (TM) quotas in an Fe-deficient Antarctic diatom. Decreases in both oxygen evolution (through photosynthesis, P) and uptake (respiration, R) coincided with a lowered growth rate of Fe-deficient cells. In addition, cells displayed reduced electron transport rates (ETR) and chlorophyll a (Chla) content, resulting in reduced cellular POC formation. Interestingly, no differences were observed in non-photochemical quenching (NPQ) or in the ratio of gross photosynthesis to respiration (GP:R). Furthermore, TM quotas were measured, which represent an important and rarely quantified parameter in previous studies. Cellular quotas of manganese, zinc, cobalt and copper remained unchanged while Fe quotas of Fe-deficient cells were reduced by 60% compared with High Fe cells. Based on our data, Fe-deficient Chaetoceros cf. simplex cells were able to efficiently acclimate to low Fe conditions, reducing their intracellular Fe concentrations, the number of functional reaction centers of photosystem II (RCII) and photosynthetic rates, thus avoiding light absorption rather than dissipating the energy through NPQ. Our results demonstrate how Chaetoceros cf. simplex can adapt their physiology to lowered assimilatory metabolism by decreasing respiratory losses.