Efforts to advance Autonomous Vehicles (AVs) have taken on a central role in research and development in recent years and will have a significant influence on road traffic in the future. Research on AVs has mainly focused on the technology itself and the direct users of AVs and their acceptance. However, the role of bicyclists, interacting with AVs in traffic, is not yet researched as thoroughly. Using a mixed methods approach, we combine quantitative results from a survey among bicyclists (N = 889) and qualitative results from a focus group (N = 19) to give insights into bicyclists’ attitudes and expectations towards self-driving cars. The results showed that bicyclists’ affinity for technology is a significant predictor for both their trust and perceived safety towards self-driving cars, as well as an effect of age and gender on these variables. Both from the quantitative and qualitative results, it is clear that flawless functioning of the technology of AVs is a prerequisite for bicyclists encountering and interacting with AVs in traffic, and that the status of the vehicle (autonomous vs. non-autonomous) is very important as well as easy to understand signals that indicate the next manoeuvres of the AV. For supporting interaction with AVs, we found that bicyclists are open to External Human Machine Interface (eHMI) solutions, as long as these ensure inclusion and support the easily-accessible nature of bicycling. Our findings can inform the design of eHMIs that help shape the interaction between bicyclists and AVs in the future, and provide insights on which factors determine the perception of AVs and, ultimately, the acceptance of AVs as part of road traffic.