This paper describes a new software package created to enhance precision during robot-assisted laser phonomicrosurgery procedures. The new software is composed of three tools for camera calibration, automatic tumor segmentation, and laser tracking. These were designed and developed to improve the outcome of this demanding microsurgical technique, and were tested herein to produce quantitative performance data. The experimental setup was based on the motorized laser micromanipulator created by Istituto Italiano di Tecnologia and the experimental protocols followed are fully described in this paper. The results show the new tools are robust and effective: The camera calibration tool reduced residual errors (RMSE) to 0.009 ± 0.002 mm under 40× microscope magnification; the automatic tumor segmentation tool resulted in deep lesion segmentations comparable to manual segmentations (RMSE= 0.160 ± 0.028 mm under 40× magnification); and the laser tracker tool proved to be reliable even during cutting procedures (RMSE= 0.073 ± 0.023 mm under 40× magnification). These results demonstrate the new software package can provide excellent improvements to the previous microsurgical system, leading to important enhancements in surgical outcome.