The fractional volume occupied by lipofuscin granules in epithelial cells of the midgut or oenocytes of abdominal fat body of 3-day-old and 13-day-old male houseflies was determined in two groups of flies by electron microscopic morphometry. One group had developed from larvae reared on diets containing no added polyunsaturated fatty acids and the second from larvae reared on diets containing added linoleic acid. No polyunsaturated fatty acids could be detected in the lipids of the first group of flies using a method which would have detected their presence in amounts greater than 0.1% of the total esterified fatty acids. The second group contained at least two hundred times more than this minimal level. The volume of lipofuscin granules increased significantly (p less than 0.01) (about threefold for the fat body and twofold in midgut cells) between 3 days and 13 days of age but no statistically significant difference was seen between the two groups of flies at the same age. The results show that if lipofuscin formation depends on the oxidation of polyunsaturated fatty acids in the housefly, then extremely small amounts of the acids are involved which lie below the detection limit of the methods employed. The age-associated small increase of extractable fluorescence seen previously in the linoleic acid group of flies is not associated with an increase in the lipofuscin granules.