Background and PurposeDiffusion MRI tractography enables to investigate white matter pathways noninvasively by reconstructing estimated fiber pathways. However, such tractograms remain biased and nonquantitative. Several techniques have been proposed to reestablish the link between tractography and tissue microstructure by modeling the diffusion signal or fiber orientation distribution (FOD) with the given tractogram and optimizing each fiber or compartment contribution according to the diffusion signal or FOD. Nevertheless, deriving a reliable quantification of connectivity strength between different brain areas is still a challenge. Moreover, evaluating the quality of a tractogram and measuring the possible error sources contained in a specific reconstructed fiber bundle also remains difficult. Lastly, all of these optimization techniques fail if specific fiber populations within a tractogram are underrepresented, for example, due to algorithmic constraints, anatomical properties, fiber geometry or seeding patterns.MethodsIn this work, we propose an approach which enables the inspection of the quality of a tractogram optimization by evaluating the residual error signal and its FOD representation. The automated fiber quantification (AFQ) is applied, whereby the framework is extended to reflect not only scalar diffusion metrics along a fiber bundle, but also directionally dependent FOD amplitudes along and perpendicular to the fiber direction. Furthermore, we also present an up‐sampling procedure to increase the number of streamlines of a given fiber population. The introduced error metrics and fiber up‐sampling method are tested and evaluated on single‐shell diffusion data sets of 16 healthy volunteers.Results and ConclusionAnalyzing the introduced error measures on specific fiber bundles shows a considerable improvement in applying the up‐sampling method. Additionally, the error metrics provide a useful tool to spot and identify potential error sources in tractograms.