Laser micromachining has emerged as a promising technique for mass production of microfluidic devices. However, control and optimization of process parameters, and design of substrate materials are still ongoing challenges for the widespread application of laser micromachining. This article reports a systematic study on the effect of laser system parameters and thermo-physical properties of substrate materials on laser micromachining. Three dimensional transient heat conduction equation with a Gaussian laser heat source was solved using finite element based Multiphysics software COMSOL 5.2a. Large heat convection coefficients were used to consider the rapid phase transition of the material during the laser treatment. The depth of the laser cut was measured by removing material at a pre-set temperature. The grid independent analysis was performed for ensuring the accuracy of the model. The results show that laser power and scanning speed have a strong effect on the channel depth, while the level of focus of the laser beam contributes in determining both the depth and width of the channel. Higher thermal conductivity results deeper in cuts, in contrast the higher specific heat produces shallower channels for a given condition. These findings can help in designing and optimizing process parameters for laser micromachining of microfluidic devices.