Relative (stratigraphy, technomorphological analysis of lithics, identification of micro-fossils, palynology) and absolute dating methods (pIRIR, palaeomagnetism) have been used in combination to establish the chronology of the Lower Palaeolithic settlement on Lesbos. Four samples from the Rodafnidia trenches have been analysed at the National Centre of Scientific Research DEMOKRITOS, Athens, using the post infrared stimulated luminescence pIRIR method, and the Thiel et al. (2011) protocol, and published in Galanidou et al., 2016. According to the resulting dates, the fluvio-lacustrine depositional units where the stone tools are excavated were last exposed to solar radiation between 476 and 164 thousand years ago, before they were covered by other deposits. Consequently, these dates provide a terminus ante quem for the activity of hominin groups in southwestern Lesbos. The chronology is in agreement with the technological and typological features of the archaeological finds.