It is currently admitted that for each filtration process using pleated filters, at least three steps can be distinguished: depth and surface filtration, which are common to flat filters, and surface reduction. This step is caused by inefficient filling of the pleat due to the filter geometry. For combustion aerosol, it has been proved that this third step strongly depends on the filtration velocity resulting in an increase of the resistance when air flow decreases. This observation leads one to think that Brownian diffusion, higher for low velocities, could influence the clogging dynamic of a pleated filter. In this article, a protocol derived from the dust cake preparation method published by Schmidt is developed. The aim of this study is to measure the aerosol penetration inside a filter media as well as in a pleat using a scanning electronic microscope and energy dispersive X-ray spectroscopy elementary detection. This method has also been extended to the study of pleated filters to measure the particle distribution inside the pleat. Filters were loaded with nanoparticles in order to evaluate the specificity of the diffusional regime on the clogging of pleated HEPA filters. For pleated filters, two filtration velocities were investigated: 2.5 and 0.2 cm/s.