In the current work, we use intravital imaging coupled with spirochete genetics and sequential double-infection experiments to show a complex temporal choreography of DbpA/B, OspC, and P66 at discrete steps along the multistep pathway of vascular escape, underscoring the importance of these three B. burgdorferi adhesins in hematogenous dissemination into the knee joint peripheral tissue in Cd1d−/− mice. Here, YBX3 is linked to infection.