Mechanical ventilation of the lung could affect surfactant turnover by alteration of its secretion, recycling, and degradation. In vitro studies of surfactant subfractions recoverable from lavage fluid have led to predictions about surfactant physiology in vivo that include morphological transformations. We used electron microscopy to study in situ lipid forms in alveoli of rat lungs after two ventilation strategies [15 min at pressures (cmH₂O) of 20/0 or 20/10]. In control animals, 4% of the lipid profile area in the surface lining layer was myelin figures (MF), 14% was tubular myelin, 37% was vesicular forms (VF), and the remainder (45%) was hypophase. Compared with controls, the length-normalized sum of the lipid forms and the hypophase was two times as great in the lungs of the 20/0 group. MF were threefold higher in the 20/0 group and fivefold higher in the 20/10 group. VF doubled after ventilation at 20/0, but VF were the same as control after ventilation at 20/10. The results showed that a ventilation pattern of 20/0 compared with that of 20/10 group was associated with a significantly larger VF, suggesting an increased net production of these surfactant forms during a large tidal volume breathing pattern. These morphological results are consistent with published results using physical methods of fractionating lung lavage.