Abstract We investigated the effect of dietary vitamin E on lipofuscin accumulation with age in the hippocampus, the inferior olive and the cerebellum of young (3–5 months old) middle-aged (12–14 months old) and old (24–26 months old) male Sprague-Dawley rats. The rats were fed either a vitamin E-deficient diet, vitamin E-supplemented diet or a control after reacbing four weeks old. We employed both quantitative light microscopy using semithin sections and qualitative fluorescence microscopy for the analysis of lipofuscin accumulation with age. The concentrations of α-tocopherol were measured simultaneously in both the plasma and the three brain regions investigated. The effect of vitamin E deficiency was statistically significant only in the inferior olive of young rats and in all the three brain regions of middle-aged rats. The effect of vitamins E supplementation was statistically significant in all three brain regions of middle-aged rats. There was no statistically significant effect of vitamin E deficiency of supplementation on lipofuscin accumulation with age as compared with the control rats in all three brain regions of old rats. It was thus revealed that dietary vitamin E clearly had a significant effect on lipofuscin and accumulation with age in the rat brain up until middle age, and that the same effect became indistinct in the latter half of their life.