The surface of a part can have an essential influence on its dynamic load bearing capacity as grooves and scratches act as more or less significant imperfections or additional micro-notches, depending on the interpretation. In order to accurately predict the fatigue strength, detailed knowledge of the surface structure in critical areas is essential but cannot always be established due to geometric constraints. To overcome these restrictions, a method utilizing impressions and a laser scanning microscope to take surface topography measurements in critical notch areas is used in this work. It enables precise and reproducible measurements to be taken. The method is used to conduct surface roughness measurements on fatigue strength specimens made of four different materials. Several roughness influence factors are calculated based on the results and their influence on the result of a recalculation using a fatigue strength assessment is shown as example. The calculated fatigue limit is compared against experimental results. The comparison shows significant scatter in prediction accuracy and no optimal surface influence factor can be recommended so far. It has to be noted that only a small sample of specimens, which were not designed for researching surface roughness influence in particular, were available for this work. The effects of roughness in the notch may not be completely captured by the available roughness influence factors. Furthermore, issues arise in the correct implementation of stress concentration factor based roughness influence factors. Further research is required to provide a more comprehensive understanding of the effects of surface roughness on the fatigue limit.