Lascaux Cave is renowned for its outstanding prehistoric paintings, strikingly well-preserved over about 18,000 yr. While stalagmites and stalactites are almost absent in the cave, there is an extensive calcite flowstone that covered a large part of the cave until its opening for tourists during the 1950s. The deposit comprises a succession of calcite rims, or “gours,” which allowed seepage water to pond in large areas in the cave. Their possible role in preservation of the cave paintings has often been evoked, but until now this deposit has not been studied in detail. Here, we present 24 new radiocarbon accelerator mass spectrometry (AMS) and 6 uranium-thorium (U-Th) analyses from the calcite of the gours, 4 AMS 14C dates from charcoals trapped in the calcite, and 4 AMS 14C analyses on organic matter extracted from the calcite. Combining the calibrated 14C ages obtained on charcoals and organic matter and U-Th ages from 14C analyses made on the carbonate, has allowed the calculation of the dead carbon proportion (dcp) of the carbonate deposits. The latter, used with the initial atmospheric 14C activities reconstructed with the new IntCal09 calibration data, allows high-resolution age estimation of the gour calcite samples and their growth rates. The carbonate deposit grew between 9530 and 6635 yr cal BP (for dcp = 10.7 ± 1.8%; 2 σ) or between 8518 and 5489 yr cal BP (for dcp = 20.5 ± 1.9%; 2 σ). This coincides with humid periods that can be related to the Atlantic period in Europe and to Sapropel 1 in the eastern Mediterranean Sea. However, geomorphological changes at the cave entrance might also have played a role in the gour development. In the 1940s, when humans entered the cave for the first time since its prehistoric occupation, the calcite gours had already been inactive for several thousand years.