Numerous studies have previously evaluated the marginal and internal fit of fixed prostheses; however, few reports have performed an objective comparison of the various methods used for their assessment. The purpose of this study was to compare five marginal and internal fit assessment methods for fixed prostheses. A specially designed sample was used to measure the marginal and internal fit of the prosthesis according to the cross-sectional method (CSM), silicone replica technique (SRT), triple scan method (TSM), micro-computed tomography (MCT), and optical coherence tomography (OCT). The five methods showed significant differences in the four regions that were assessed (p < 0.001). The marginal, axial, angle, and occlusal regions showed low mean values: CSM (23.2 µm), TSM (56.3 µm), MCT (84.3 µm), and MCT (102.6 µm), respectively. The marginal fit for each method was in the range of 23.2–83.4 µm and internal fit (axial, angle, and occlusal) ranged from 44.8–95.9 µm, 84.3–128.6 µm, and 102.6–140.5 µm, respectively. The marginal and internal fit showed significant differences depending on the method. Even if the assessment values of the marginal and internal fit are found to be in the allowable clinical range, the differences in the values according to the method should be considered.