Therapeutic drug carriers can drive their cargo to their target cells. However, an obstacle is usually the entrapment of the drug inside the endolysosomal compartment, which physically impedes its actuation by the impossibility of reaching its molecular site of action. To overcome this hurdle, photochemical internalization (PCI) has been proposed, but the extent of PCI-induced membrane disruption and its capability to allow the release of microparticles is unknown. The aim of the present study was to determine if PCI allows the release of microparticles from the endolysosomal compartment to the cytosol and to analyze at the ultrastructural level the effect of PCI on the membrane surrounding the particles. Confocal microscope allowed us to detect that endolysosomal membranes suffered some disruption after PCI, evidenced by the diffusion of soluble transferrin from the endolysosomes to the cytosol and by a decrease of LAMP1-microparticles co-localization. Transmission electron microscopy (TEM) showed a decrease in the number of well-defined membranes around microparticles after PCI, and scanning TEM combined with energy dispersive x-ray revealed an increase in the width of endolysosomal membranes after treatment. These results suggest that endolysosomal membranes suffered an ultrastructure alteration after PCI, enough to liberate soluble transferrin but not the entire microparticles.