Measuring food intake in a laboratory usually involves limited food choices. An automated food-selection system with two vending machines containing a large variety of foods was used to measure food intake in 10 male volunteers (31 +/- 6 y, 69.2 +/- 7.1 kg, 18 +/- 7% fat, mean +/- SD) on a metabolic ward. The effect of carbohydrate, fat, and protein intakes on 24-h energy expenditure (24EE) and substrate oxidations was measured in a respiratory chamber during day 4 of weight maintenance and day 7 of ad libitum intake. Ad libitum intake resulted in a 7-d overfeeding of 6468 +/- 3824 kJ/d above weight-maintenance requirements, leading to a 2.3 +/- 1.2-kg gain. The 10,975 +/- 3774 kJ excess energy intake on day 7 of ad libitum intake caused a 1205 +/- 920 kJ/d increase in 24EE (delta 24EE = 0.17 x delta intake - 695; r = 0.71, P less than 0.02). Of the excess carbohydrate intake, 74% was oxidized (r = 0.86, P less than 0.001), whereas excess fat intake was not. Carbohydrate and protein stores are regulated whereas excess fat intake is channeled to fat stores.