A method for calibrating a laser profiling system for seafloor micro-topography measurements is described. The system consists of a digital camera and an arrangement of six red lasers that are mounted as a unit on a remotely operated vehicle (ROV). The lasers project as parallel planes onto the seafloor, creating profiles of the local topography that are interpreted from the digital camera image. The goal of the calibration was to determine the plane equations for the six lasers relative to the camera. This was accomplished in two stages. First, distortions in the digital image were corrected using an interpolation method based on a virtual pinhole camera model. Then, the laser planes were determined according to their intersections with a calibration target. The position and orientation of the target were obtained by a registration process. The selection of the target shape and size was found to be critical to a successful calibration at sea, due to the limitations in the manoeuvrability of the ROV.