The invention discloses an image signal generation method and device with a coordinate acquisition function, and relates to the technical field of display panel detection. The method includes the steps: S1) acquiring vertical line position indication signals and horizontal line position indication signals according to resolution ratio configuration information and (X, Y) coordinate configuration information; S2) acquiring cross line coordinate point information according to the vertical line position indication signals and the horizontal line position indication signals, and configuring RGB (red green blue) values of a cross line coordinate point; S3) correspondingly distributing the RGB values of the cross line coordinate point to RGB image data of links to obtain cross line image signals. The cross line image signals with real-time configurable color and center point coordinates are generated, an optical calibrator can be assisted in visually and rapidly acquiring coordinate information of defect points or defect areas of a display panel to be detected, and a laser repair machine can be assisted in improving the bright spot and dark spot repair efficiency of the display panel.