Neurodevelopmental disorders (NDDs) are often associated with mutations in mitotic genes rather than in genes specifying neuronal functions. Several mitotic regulators are functionally repurposed to new roles during neurodifferentiation. CENP-F, a kinetochore- and microtubule-interacting protein with well-characterised mitotic functions, is being growingly identified for harbouring mutations in primary microcephaly and in the Strømme syndrome, an NDD classified as a ciliopathy. The mechanism through which mutant CENP-F contributes to these NDDs remains elusive.To gain insight into the implication of CENP-F in NDDs, we have optimised imaging methods and developed a comprehensive analysis of CENP-F in post-mitotic cellular systems. We first examined CENP-F during the formation of the primary cilium by optimising quantitative immunofluorescence and single-cell analysis of spinning-disk confocal images. CENP-F and some of its interacting partners were characterised in a time-course analysis of ciliogenesis in human retinal RPE-1 cells. CENP-F localises at the basal body of the cilium and is absolutely required for proper ciliogenesis. We also studied the requirement for CENP-F during neurodifferentiation by time-lapse videorecording and confocal microscopy. We have developed two AI-based algorithms to analyse high-content images of videorecorded cell cultures: that enabled us to extract information on the dynamics with which phenotypic changes occur during neurodifferentiation and identify steps that are sensitive to and/or impaired by CENP-F loss-of-function. In summary, these imaging protocols identify critical temporal windows during which the lack of CENP-F impacts neurodifferentiation and ciliogenesis, thus providing new tools to dissect roles of CENP-F in NDDs.