Determination of the internalization pathway of photoluminescent nanodiamonds in mammalian cells for biological labeling and optimization of the fluorescent yield.
Résumé
Diamond nanoparticles have been recently used as new fluorescent labels in cells. Their emission relies on color centers embedded in the diamond matrix. In this work we compare the photoluminescence of a single color center embedded in a 30 nm diameter nanodiamond to that to a single dye molecule and demonstrate the perfect photostability of the color centers. We also compare the photoluminescence properties of nanodiamonds prepared under different conditions in order to find the optimal parameters to achieve a high fluorescence yield. We use these photoluminescent nanodiamonds for HeLa cell labeling and investigate their uptake mechanism. On the one hand by immunostaining the endocytotic vesicles and on the other by transmission electron microscopy observations we study the localization of nanodiamonds in cells. Moreover, by blocking selectively different endocytotic mechanisms we unravel their internalization pathway. We find that nanodiamonds enter the cells by receptor-mediated endocytosis. The results of this study are a prerequisite for the use of photoluminescent nanodiamonds as intracellular markers or drug delivery devices.
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