Interfacial adhesion is a major concern with respect to successful performance of thin polymer films in developing new thin-film processes. Micro-indentation was used to induce interfacial delamination of polytetrafluoroethylene (PTFE) films deposited on glass substrates using hot filament chemical vapour deposition (HFCVD). Film thickness (1, 2, 3, 5, 10 mu m) and indentation load (0.5, 0.75, 1, 2, 3 N) effects on the delamination diameter were investigated. A three-dimensional finite element model using shear material failure criterion and cohesive zone model (CZM) was developed to simulate the delamination. A normalized load-delamination radius relationship was obtained to evaluate the interfacial fracture toughness. The experimental observations showed that the delamination diameter depends on film thickness and indentation load. The numerical simulation indicates the delamination diameter depends on film thickness, material properties, and indentation force. The predictions of interfacial fracture toughness for 5- and 10-mu m PTFE films are much smaller than those values using Rosenfeld et al.'s equation, which excludes the energy spent during the penetration.