Teflon, polytetrafluorethylene (PTFE), is an important material in bioscience and medical application due to its special characteristics (non-flammable, anti-adhesive, heat-resistant and bio-compatible). The advantages of ultrashort laser processing of Teflon include a minimal thermal penetration region and low processing temperatures, precision removal of material, and good-quality feature definition. In this paper, laser processing of Teflon by Ti:Sapphire femtosecond laser (780 nm, 110 fs), Nd:YAG laser (532 nm, 7 ns) and CO2 laser (10.6 μm, 10 μs) has been investigated. For femtosecond laser processing, clear ablation takes place and provides high-quality groove on Teflon surface. Both the groove depth and the width increase as the laser fluence increase, and decrease almost linearly as the scanning speed increase for laser fluence below 5.0 J/cm2. For Nd:YAG processing, Teflon surface roughness is improved but no clean ablation is accessible, which makes it difficult to micromachine Teflon by Nd:YAG laser. For CO2 laser processing, laser-induced bumps were formed on Teflon surface with controlled laser parameters. The physics mechanisms for different pulse duration laser processing of Teflon are also discussed.