The internal fouling of membranes is typically presumed and inferred, but the direct characterization of representative samples is challenging. This study targeted to assess internal fouling using the Optical Coherence Tomography (OCT) and Evapoporometry (EP) techniques for real-time monitoring during filtration and off-line measurement of the pore size distributions (PSDs) of the fouled membranes, respectively. The results were validated by atomic force microscopy (AFM) measurements and also the three-mechanism fouling model. The foulant used was the well-studied bovine serum albumin (BSA), while the membranes were three commercially available polymeric microfiltration membranes with similar nominal pore sizes and porosities. Although the OCT affirmed the progressive worsening of internal fouling in real-time, the quantitative comparison of the extents of internal fouling among the three membranes was not possible. As for EP, it was able to quantitatively compare the pore size distributions and average pore diameters to ascertain the different extents of internal fouling. The phenomenological model available was effective in quantitatively comparing the extents of pore-constriction among the three membranes, while AFM tied the worst internal fouling to the most attractive BSA-membrane affinity. More in-depth understanding of internal fouling is warranted to not only recommend better membranes but also facilitate the advancement of models.