Humans are unable to synthesize linoleic acid (LA) (18:2 omega-6) and alpha-linolenic acid (LNA) (18:3 omega-3). Most formulas provide ample LA, yet infants are at risk for omega-3 deficiency unless they are fed human milk. Neonates born at 30 weeks received human milk or were randomized to three formulas: formula A, based on corn oil, similar to old commercial formula; formula B, based on soy oil supplied LNA; or formula C, a product similar to B with added marine oil to provide docosahexaenoic acid (22:6 omega-3). The fatty acids of plasma and red blood cells had marked diet-induced differences. The rod photo-receptor tests demonstrated higher threshold and decreased sensitivity in the omega-3-deficient infants. Visual acuity also showed improved function of the brain cortex in the human milk and group C infants at follow-up at 57 weeks. These results suggest that omega-3 fatty acids are needed for optimal development of visual function.