AbstractAlveolar epithelial type I (AE1) cells with their wide spatial expansion form approximately 95% of the outer surface area of the air-blood barrier inside the lung. Serial block-face scanning electron microscopy (SBF-SEM) investigations led to the hypothesis that AE1 cell mitochondria are preferentially distributed as aggregates in those parts of AE1 cells that are located above connective tissue pillars between capillaries, thus not increasing the thickness of the diffusion distance for oxygen and carbon dioxide. Furthermore, it was hypothesised that postnatal development requires adapting the amount and distribution of mitochondria in AE1 cells. Human lung samples from three infant (26 and 30 days, 6 months) and three adult (20, 39 and 40 years) samples were investigated by light microscopy, transmission electron microscopy and stereology. The volume fraction of mitochondria was similar in infant and adult lungs with a mean value of 6.3%. The ratio between mitochondrial profiles on top of capillaries or above connective tissue pillars was approximately 3:1 in infants and adults. However, regarding the volume of both cytoplasmic compartments, infants showed a higher number of mitochondrial profiles on top of capillaries while adults showed a higher number above connective tissue pillars. Samples of three additional adult lungs were analysed by confocal laser scanning microscopy. Again, mitochondria were not preferentially found as aggregates above connective tissue pillars. In conclusion, AE1 cell mitochondria were not preferentially found as aggregates, showed the same volume density in infants and adults but differed in distribution between the age groups.