A model system for multicompartment nanofibers was fabricated by colloid electrospinning. The obtained nanostructured material consisted of fluorescent polymer nanoparticles that were synthesized in a miniemulsion and then embedded in fluorescently labeled polymer nanofibers. Because of the absence of contrast between both polymers, the immobilized nanoparticles cannot be reliably identified in the nanofibers via electron microscopy or other techniques. Here, we describe investigations on the hybrid material with two-color STED microscopy to localize the nanoparticles and to quantify their distribution along nanofibers with particle and fiber radii down to 50 nm.