Abstract Some of the most remarkable Neolithic finds encountered in central Europe are wells with a wooden construction. These features provide unusual insights into Neolithic societies, their subsistence strategies and the landscape they inhabited. In the last fifteen years, four Early Neolithic wells out of a total of seven such wells known since the 1970s have been discovered in Czechia. One of them is the well at Ostrov, the wooden construction and sedimentary infill of which have made it possible to study diverse topics by taking a multi-proxy approach using a range of complementary methods. The study presented here compares different periods of the well’s existence, examines the carpentry skills of the first farmers, investigates sedimentary processes, reveals details about the use and decline of the well, addresses its location relative to that of the settlement, and reconstructs the environment of the well itself and the surrounding landscape. The water well at Ostrov is a highly unusual find because it probably existed outside of a settlement. The grooved wooden lining used is identical to that of previously investigated wells inside settlements, as are the processes of its use and decline. However, environment-related indicates that the Ostrov well was located in an open landscape with sparse vegetation and was perhaps only used by-passing herdsmen.