The role of selenium in human brain physiology, as well as in aging and neurodegenerative processes, remains unclear. Thus, the aim of this study was to establish the "normal" (reference) levels for selenium in the human brain, as well as anatomical regional differences and age-related changes. Using inductively coupled plasma-mass spectrometry after sample microwave-assisted acid digestion, selenium levels were measured in 14 different areas of the brain of adult individuals (n = 42; 71 ± 12, range 50-101 years old) without a known history of neurodegenerative, neurological, or psychiatric disorders. In the whole data set (n = 588; 42 individuals × 14 brain areas), selenium levels ranged from 552 to 1435 ng/g, with a mean ± SD content of 959 ± 178 ng/g (dry weight basis). Selenium distribution across the different brain areas was heterogeneous, with the highest levels in the putamen, parietal inferior lobule, and occipital cortex and the lowest expression in the medulla and cerebellum. Selenium levels were unchanged with aging. Compared with the age-matched control group, significantly increased levels of selenium were found in the globus pallidus, superior temporal gyrus, and frontal cortex of Parkinson's disease (n = 1) and Alzheimer's disease (n = 2) patients. This study provides new data on the anatomical regional differences in selenium levels in the human brain, which will aid future interpretation of studies examining brain tissue affected by neurodegenerative (and other) brain diseases.