AbstractDiffusion MRI of the brain enables to quantify white matter fiber orientations noninvasively. Several approaches have been proposed to estimate such characteristics from diffusion MRI data with spherical deconvolution being one of the most widely used methods. Constrained spherical deconvolution requires to define – or derive from the data – a response function, which is used to compute the fiber orientation distribution (FOD). This definition or derivation is not unequivocal and can thus result in different characteristics of the response function which are expected to affect the FOD computation and the subsequent fiber tracking. In this work, we explored the effects of inaccuracies in the shape and scaling factors of the response function on the FOD characteristics. With simulations, we show that underestimation of the shape factor in the response functions has a larger effect on the FOD peaks than overestimation of the shape factor, whereas the latter will cause more spurious peaks. Moreover, crossing fiber populations with a smaller separation angle were more sensitive to the response function inaccuracy than fiber populations with more orthogonal separation angles. Furthermore, the FOD characteristics show deviations as a result of modified shape and scaling factors of the response function. Results with the in vivo data demonstrate that the deviations of the FODs and spurious peaks can further deviate the termination of propagation in fiber tracking. This work highlights the importance of proper definition of the response function and how specific calibration factors can affect the FOD and fiber tractography results.