The installation pose error of the laser probe mounted on a three-axis CNC machine tool is not easy to adjust, which may result in measurement error. In this article, a posture calibration method of the laser probe for on-machine measurement (OMM) is proposed. A mathematical model based on the laser line profiler for OMM is formulated, the spatial pose of the calibration reference point is measured by the laser probe, and the linear algorithm to obtain the pose parameters of the probe installation is derived based on the handeye calibration. Considering the influence of the positioning errors of the machine tool on the accuracy of the calibration, an error analysis is implemented by using Monte Carlo simulation. Experimental results show that when measuring a circular hole with a radius of 35 mm, the measurement error of the circular hole after calibration is 0. 051 6 mm, and the measurement accuracy is improved by approximately 96% , which validate the effectiveness and feasibility of the proposed calibration method.