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Abstract Since wound healing requires cells to repopulate a damaged area, cell migration is essential. In addition, migration plays a crucial role in cancer metastasis. Whether tumour cells can invade tissue and metastasize is a crucial factor determining their malignancy or in other words a hallmark of cancer (Lazebnik in Nat Rev Cancer 10: 232–233, 2010, https://doi.org/10.1038/nrc2827 ). Nanodiamonds potentially offer a powerful tool to investigate these migration processes. Due to their unprecedented photostability, they can function as long-term fluorescent labels. Besides, nanodiamonds are robust quantum sensors that can reveal, for instance, the temperature or the concentration of certain chemicals with nanoscale resolution. However, to utilise nanodiamonds to study cell migration, it is essential to understand if and how the presence of nanodiamonds influences cell migration. Here, we investigate this process for the first time. We found that nanodiamonds do not alter the speed at which HeLa cells populate a scratch at any tested concentrations. Furthermore, we tested cell attachment by quantifying focal adhesion points. Oxygen-terminated fluorescent nanodiamonds influence the cell spreading, the number of focal adhesions and the size of focal adhesion points. Interestingly, this is different for other types of nanodiamonds in the literature. For these particles, it has been described in the literature that they hinder cell migration. Our results support that fluorescent nanodiamonds do not influence cell migration strongly and thus can be used in labelling and sensing migrating cells. Graphical Abstract