The rapid increase of e-bike use in the Netherlands has intensified speed heterogeneity on cycling infrastructure. Although this trend contributes positively to sustainable mobility, it may negatively influence cyclists’ perceived safety, which in turn affects their route choices and willingness to cycle. Existing literature recognizes the role of infrastructural, environmental, traffic-related, and individual factors in shaping perceived safety, yet the specific influence of speed differences in bicycle-only environments remains insufficiently studied. This research investigates the extent to which speed heterogeneity affects perceived cycling safety and how this effect interacts with other contextual attributes. A video-based stated preference experiment was designed in which respondents evaluated overtaking events filmed in real urban locations, manipulated across lane design, lane width, cycling density, and speed differences. A panel Mixed Logit model was estimated, supplemented with scenario ranking tasks. Results show that greater speed differences consistently reduce
perceived safety, especially under congested conditions. Two-way lanes are perceived as less safe than one-way lanes, though wider lanes mitigate this effect and experienced cyclists express higher comfort levels. Findings underline the importance of managing
speed heterogeneity through infrastructural design and policy interventions, including multi-paced cycling lanes, speed awareness
campaigns, and targeted safety reporting mechanisms. The study demonstrates the value of video-based stated preference methods for analyzing subjective safety and contributes to policy debates on designing cycling infrastructure for increasingly diverse urban cyclists.