The effects of various fixative-buffer combinations on lung fine structure.

A variety of fixatives, buffers and fixation procedures were compared in rat and squirrel monkey lung in an attempt to preserve optimally both the cytologic details of pulmonary parenchyma as well as the acellular avelolar lining layer. In initial experiments utilizing the fixative of Ito and Karnovsky ('68), an electron-dense deposit was observed on the alveolar surface. Experiments were carried out in an attempt to determine what component of this fixative was responsible for the reaction product observed. In addition,immersion fixation of tissue blocks was compared to the whole lung fixation method of Kikkawa ('70). Kikkawa ('70) achieved excellent preservation of the acellular alveolar lining layer by such a fixation technique. In all lungs examined, whenever a phosphate buffer was utilized with primary aldehyde fixation, an electron-dense precipitate was observed on the luminal surfaces of the type I and II pulmonary epithelial cells. Additional sites of reaction product were pinocytotic vesicles of the type I cells and membranous arrays within the alveolar lumen. Such deposits were never observed when a sodium cacodylate buffer was used. No such granules were observed in areas of lung where the acellular alveolar lining layer had been preserved. The implications of these findings with regard to lung histochemical procedures and the possible relationship of these phosphate buffer-dependent granules to the surfactant system are discussed.