To study interactions between Ixodes scapularis (Say) and Borrelia burgdorferi, an artificial feeding system was refined to allow controlled manipulation of single variables. The feeding system uses a mouse skin mounted on a water-jacketed glass membrane feeder. I. scapularis were infected using either BSK-H-cultured B. burgdorferi spirochetes or a B. burgdorferi-infected mouse skin as the source of spirochetes. Sixty-six percent of nymphs successfully fed to repletion using the artificial feeding systems with at least 75% of nymphs becoming infected with B. burgdorferi. Strain B31 B. burgdorferi spirochetes from passages 2–17 were equally infectious to nymphal ticks. At concentrations of one spirochete per microliter, 12% of nymphs acquired infection and 14 and 100 spirochetes per microliter resulted in 50 and 100% infection rates, respectively. Eighty-nine percent of nymphs fed by artificial feeding molted to the adult stage. When subsequently fed as adults, these I. scapularis successfully transmitted infectious B. burgdorferi spirochetes to mice.