The three-dimensional (3D) spatial structure of an iron oxide containing alkyd paint specimen has been investigated by serial block-face scanning electron microscopy (SBFSEM). The resultant images of the 3D structure clearly present the spatial distribution of the iron oxide pigment particles in the coating film and reveal the extent of aggregation of the particles in the matrix material. More than one-half of the iron oxide particles (in volume) had aggregated to form clusters of considerable sizes that follow a Gaussian spacing distribution in the measured coating film. Over 80% of the clusters have dimensions between 1.5μm and 3.5μm; also, pores are evident at the centres of clusters whose sizes are larger than 2μm. The work demonstrated here reveals a new approach to fully characterize the 3D spatial structure of coatings and to explore their correlations with the performance of the materials.