Systematic dating of tufa samples has been undertaken to establish a chronology of tufa deposits in Plitvice National Park, Yugoslavia. We conclude that tufa samples give reproducible data within the time span of ≈ 40,000 years up to recent. The 14C/12C ratio of carbon in the ambient biosphere and hydrosphere gives a detailed picture of the distribution of carbon isotopes in the investigated system. Despite the susceptibility of calcareous material to ambient conditions in the hydrosphere, the original 14C composition of tufa has not been substantially changed. A vertical profile which was cut in tufa during pathway construction showed that the tufa isochrones run vertically in the investigated area. The vertical isochrones and the 14C gradient were horizontal due to tufa build-up in Plitvice National Park which occurs on vertical escarpments that have been flooded. The relatively uniform radiocarbon content of most tufa deposits indicates redistribution of radiocarbon during the formation period. While detailed stratigraphy is often obscured, a general chronology of tufa deposits in the investigated area has been established.