Iron levels in the healthy human brain are known to be high in certain areas such as the substantia nigra (SN), and increase further with age. In addition, there is some evidence for a further increase in iron load in the SN of Parkinsons disease (PD) patients as compared to controls, which correlates with motor disability. Here, we have analyzed total iron levels in cells as well as mouse and human brain samples by atomic absorption spectroscopy (AAS). Our data indicate that iron load is more pronounced in cells with dopaminergic features. Moreover, region-specific differences in iron load reflecting those in the human brain were detected in rodent brains as well. Whilst altered iron load was not observed in other regions also affected in PD patients, we report a significant increase in iron load in the SN of Lewy body disease patients as compared to Alzheimers disease (AD) patients or controls, which correlates with neurodegeneration in this brain area.