Tissue engineering (TE) strategies are based on the active interplay among cells, three-dimensional (3D) scaffolds, and physiological signals in view of developing in vitro functional tissue substitutes. Scaffold effectiveness is strongly influenced by its microstructure and ability to be permeated by fluids (i.e., scaffold permeability) and consequently to be colonized by cells. Permeability depends on the combination of porosity, pore size, tortuosity, and interconnectivity and its characterization is crucial for an effective evaluation of the overall scaffold performance. Several methods and set-ups have been proposed to characterize the permeability of hard or soft materials, but no systems able to characterize both types of materials have yet been developed.