We measured the single breath diffusing capacity for carbon monoxide (DLcoSB) using a three-equation method to describe CO uptake in 10 normal seated subjects who either voluntarily inhaled slowly (0.5 L/sec) to total lung capacity (TLC), or inhaled slowly to TLC with maximal effort through a high inspiratory resistance which created high negative inspiratory pressure. Subjects then immediately exhaled slowly at a voluntarily controlled exhaled flow. Single breath maneuvers were performed in duplicate both with and without high negative inspiratory pressure while subjects were seated upright at rest and during steady-state bicycle exercise. We found that high negative inspiratory pressure increased DLcoSB by 10.5 +/- 4.9% (mean +/- 1 SD) at rest (P less than 0.001). In 7 subjects low level exercise alone increased DLcoSB by a similar amount (12.1 +/- 7.3%; P = 0.005). In six of the subjects there was a significant correlation between the increase in DLcoSB during high negative inspiratory pressure at rest and the increase in DLcoSB during steady-state exercise (r = 0.89; P less than 0.01). During steady-state exercise, high negative inspiratory pressure further increased DLcoSB 6.4 +/- 6.3% compared to exercise alone (P = 0.05). We conclude that the increase in DLcoSB with high negative inspiratory pressure at rest is a simple reproducible method of assessing recruitment of the pulmonary capillary bed in man.