A stomatal frequency record based on buried Tsuga heterophylla needles reveals significant centennial-scale atmospheric CO2 fluctuations during the last millennium. The record includes four CO2 minima of 260–275 ppmv (ca. A.D. 860 and A.D. 1150, and less prominently, ca. A.D. 1600 and 1800). Alternating CO2 maxima of 300–320 ppmv are present at A.D. 1000, A.D. 1300, and ca. A.D. 1700. These CO2 fluctuations parallel global terrestrial air temperature changes, as well as oceanic surface temperature fluctuations in the North Atlantic. The results obtained in this study corroborate the notion of a continuous coupling of the preindustrial atmospheric CO2 regime and climate.