The distribution of osmiophilic lamellae within the alveolar and bronchiolar walls of the mammalian lungs as revealed by osmium‐ethanol treatment

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Abstract
Osmiophilia of the cellular elements of the alveoli and bronchioles was investigated using the new technique of second osmication with a solution containing a mixture of osmium tetroxide and ethanol. Previously this method was used to reveal osmiophilic periodic lamellae of synthetic dipalmitoyl lecithin and the surface film of the acellular alveolar lining layer of the mammalian lung in thin sections.In the present experiments, much of the osmiophilic content of inclusions in type II epithelial cells became visible only after second osmication with osmium‐ethanol but was not visible with the initial osmium tetroxide fixation. The osmiophilic content of the inclusions showed 42 Å periodicity. These results are in accord with those obtained in the study of dipalmitoyl lecithin and the surface film, suggesting that the inclusions are the precursors of the pulmonary surfactant.Several new observations were made with this technique. These include osmiophilic lamellae within Golgi saccules, within the multivesicular bodies of type II cells, in the vesicles of type I epithelial cells, and in the basement membranes of epithelial cells. These observations are consistent with the idea that the Golgi apparatus and multivesicular bodies are closely related to the production of surfactant and that the surfactant may be disposed of by type I epithelial cells after engulfment by pinocytosis. Thereafter, the surfactant may eventually be cleared through the lymphatics.Clara cell granules behaved differently and were thought to be an unlikely site for the synthesis of the pulmonary surfactant.