This study aimed to estimate intake of individual polyunsaturated fatty acids (PUFAs), identify major dietary sources of PUFAs and estimate the proportion of individuals consuming fish among US children 12–60 months of age, by age and race and ethnicity. The study employed a cross-sectional design using US National Health and Nutrition Examination Survey data. Representative sample of US population based on selected counties. Subjects: 2496 US children aged 12–60 months. Mean daily intake of n-6 PUFAs and eicosapentaenoic acid (EPA) varied by age, with children 12–24 months of age having lower average intakes (mg or g day−1) than children 49–60 months of age and the lowest n6 : n3 ratio, upon adjustment for energy intake. Docosahexaenoic acid (DHA) intake was low (20 mg day−1) compared to typical infant intake and did not change with age. Compared to non-Hispanic white children, Mexican American children had higher DHA and arachidonic acid (AA) intake. In the previous 30 days, 53.7% of children ever consumed fish. Non-Hispanic black children were more likely than non-Hispanic white children to have consumed fish (64.0% vs. 53.0%). Results indicate low prevalence of fish intake and key n-3 PUFAs, relative to n-6 fatty acids, which suggests room for improvement in the diets of US children. More research is needed to determine how increasing dietary intakes of n-3 PUFAs like DHA could benefit child health.