Hypoxia and oxidative stress are contributing factors in cardiovascular disease, and notably changes in zinc, iron and calcium homeostasis affect cellular responses to oxygen tension and oxidative stress. Dietary zinc intake correlates inversely with subclinical carotid atherosclerosis, and serum zinc levels are reduced in patients with heart failure or undergoing cardiac surgery. Although zinc is a redox-inert metal, physiological concentrations of zinc have anti-oxidant, anti-inflammatory and anti-proliferative properties, whilst zinc deficiency or overload generates oxidative stress. Zinc has also been shown to afford protection during ischaemia reperfusion injury. We have used ICP-MS (Inductively Coupled Plasma Mass Spectrometry) to map changes in total metal content in human coronary artery endothelial cells (HCAECs) cultured long-term (at least 5 days) under hyperoxia, physiological normoxia and hypoxia (18, 5 and 1 kPa O2) and exposed to ischaemia reperfusion injury. Decreases in 66Zn and increases in 56Fe were measured as cells were adapted from 18 kPa to 1 kPa O2 levels. In parallel experiments, mobilisation of intracellular free metals during exposure to in vitro ischaemia reperfusion injury was measured using metal specific fluorescent probes. Intracellular Fe2+ was unaffected by acute hypoxia but significantly increased upon re-oxygenation. Increased Fe2+ correlated with superoxide production and was abrogated by treatment with PEG-SOD but elevated by a SOD inhibitor (ammonium tetrathiomolybdate). We are currently employing Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) to further probe the spatial distribution of Zn, Fe, Ca, Cu and Mn in cells maintained under varying ambient O2 conditions in a Scitive O2-regulated workstation. Our findings highlight the importance of characterising metal fingerprints in cultured cells and tissues which will provide novel insights for potential therapeutic interventions to limit damage in ischaemia reperfusion injury.