An internal and marginal fit between prosthesis and abutment is an important factor of the durability of dental prosthesis. In this paper, we have proposed the use of swept-source optical coherence tomography (OCT) as a novel application for non-ionized and high-resolution measurements of internal and marginal discrepancies at anatomically critical four points, such as occlusal, angle, axial, and margin during prosthesis attachment. A tooth model was fabricated by a 3D printing technique, and the dental prosthesis was designed using dental CAD software. The cross-sectional images along with intensity peak profile analysis of the sample were acquired using the OCT system for measurements of fit. The demonstrated qualitative and quantitative evaluations can be well-utilized for assessment of the internal and marginal fit of dental prosthesis.