This study was performed to observe microstructures of the rat lung, using a synchrotron radiation beam and to compare findings with histological observations. X-ray refraction images from ex-vivo ventilating rat lung were obtained with an 8 KeV monochromatic beam and 20-mum thick CsI(Tl) scintillation crystal. The visual image was magnified using a 20x microscope objective and captured using an analog CCD camera. Obtained images were compared with conventional light microscopic findings from the same tissue. Pulmonary microstructures, including alveolar ducts, alveolar sacs, alveoli, alveolar walls, and perialveolar capillary networks were clearly identified with spatial resolution of as much as 1.2 mum and had good correlation with conventional light microscopic findings. The shape of alveoli appeared more round in SR images than in the light microscopic images. The results suggest that X-ray microscopy study of the lung using synchrotron radiation demonstrates the potential for clinically relevant microstructure of lung tissue without sectioning and fixation.