The regional deposition of an inhaled aerosol of 1.0-mu m diameter fluorescent microspheres (FMS) was used to produce high-resolution maps of regional ventilation. Five anesthetized, prone, mechanically ventilated pigs received two 10-min inhalations of pairs of different FMS labels, accompanied by intravenous injection of 15.0-mu m radioactive microspheres. The lungs were air dried and cut into 1.9-cm(3) pieces, with notation of the spatial coordinates for each piece. After measurement of radioactive energy peaks, the tissue samples were soaked in 2-ethoxyethyl acetate, and fluorescent emission peaks were recorded for the wavelengths specific to each fluorescence label. The correlation of fluorescence activity between simultaneously administered inhaled FMS ranged from 0.98 to 0.99. The mean coefficient of variation for ventilation for all 10 trials (47.9 +/- 8.1%) was similar to that for perfusion (46.2 +/- 6.3%). No physiologically significant gravitational gradient of ventilation or perfusion was present in the prone animals. The strongest predictor of the magnitude of regional ventilation among all animals was regional perfusion (r = 0.77 +/- 0.13).