Little is known about the influence of accident risk and perceived safety on bicycle route choice. Do accident hot spots or points of 'perceived insecure' influence actual route choice of cyclists? To answer that question a GPS data set (~4,000 trips from ~170 study participants) and further data (infrastructure, operation, exposure, accidents reports, and survey data recording critical incidents) is used. The results reveal that a high accident risk along the route shows a significant negative but slight influence on route choice. In contrast, perceived safety does not significantly influence route choice. Other factors (e.g. existence or narrow width of cycling infrastructure, higher traffic volumes of bicycle traffic, signal controlled intersections) show significant influence and may represent perceived safety better than the used survey data.