The range-dependent overlap between the laser beam and the receiver field of view of a lidar can be determined experimentally if a pure molecular backscatter signal is measured in addition to the usually observed elastic backscatter signal, which consists of a molecular component and a particle component. Two methods, the direct determination of the overlap profile and an iterative approach, are presented and applied to a lidar measurement. The measured overlap profile accounts for actual system alignment and for all system parameters that are not explicitly known, such as actual laser beam divergence and spatial intensity distribution of the laser light.