To further validate a stochastic particle deposition model, threedimensional deposition patterns predicted by that model were compared with corresponding spatial particle deposition data obtained from SPECT measurements. In the in vivo inhalation experiments, two different polydisperse aerosols with mass median aerodynamic diameters of 1.6 µm and 6.8 µm were inhaled by 12 test subjects, using different nebulizers. Predicted and measured deposition data were compared on three different levels: (1) total lung deposition, (2) deposition per hemispherical shell, and (3) deposition per airway generation. First, experimental and theoretical total lung deposition data showed good agreement for both the fine (65 ± 9% vs. 55 ± 21%) and the coarse aerosols (55 ± 8% vs. 46 ± 4%). Second, predicted deposition per hemispherical shell also corresponded well with the experimental data, both exhibiting small deposition fractions in the inner shells and a roughly quadratic increase in the outer shells. Third, fair agreement was observed for the deposition fractions per airway generation, both experimental data and modelling predictions exhibiting relatively small deposition fractions in central bronchial airway generations, followed by a steep increase in the peripheral respiratory airways. While the overall agreement between measured SPECT data and computed deposition fractions demonstrates that SPECT data can indeed be used for model validation, the current spatial resolution of the SPECT method allows only a limited validation of model predictions at the single airway generation level.