The various factors influencing closing volume were studied by performing the single-breath N2 test on 9 healthy nonsmokers. Time of day, day of the week, and preceding volume history had no effect on either closing volume or alveolar plateau. Slow inspiratory flow resulted in larger ratio of closing volume to vital capacity, ratio of closing capacity to total lung capacity, and change in N2 concentration than fast inspiratory flow. Voluntary regulation of the expiratory flow resulted in smaller ratios of closing volume to vital capacity and closing capacity to total lung capacity than when flow was regulated by a resistance. Prolonged breath holding of the inspired O2 led to larger ratio of closing volume to vital capacity and ratio of closing capacity to total lung capacity. To obtain uniform, comparable closing volumes, it is suggested that the subject inspire slowly, control expiratory flow (preferably voluntarily), and not pause between inspiration and expiration.