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Photoluminescent diamond nanoparticles for cell labeling: study of their uptake mechanism in mammalian cells

Orestis Faklaris, Vandana Joshi, Theano Irinopoulou, Patrick Tauc, Mohamed Sennour, Hugues Girard, Celine Gesset, Jean-Charles Arnault, Alain Thorel, Jean-Paul Boudou, Patrick A. Curmi, François Treussart, ACS Nano 3, 3955-3962 (2009)

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Diamond nanoparticles (nanodiamonds) have been recently proposed as new labels for cellular imaging. For small nanodiamonds (size <40 nm) resonant laser scattering and Raman scattering cross-sections are too small to allow single nanoparticle observation. Nanodiamonds can however be rendered photoluminescent with a perfect photostability at room temperature. Such a remarkable property allows easier single-particle tracking over long time-scales. In this work we use photoluminescent nanodiamonds of size <50 nm for intracellular labeling and investigate the mechanism of their uptake by living cells . By blocking selectively different uptake processes we show that nanodiamonds enter cells mainly by endocytosis and converging data indicate that it is clathrin mediated. We also examine nanodiamonds intracellular localization in endocytic vesicles using immunofluorescence and transmission electron microscopy. We find a high degree of colocalization between vesicles and the biggest nanoparticles or aggregates, while the smallest particles appear free in the cytosol. Our results pave the way for the use of photoluminescent nanodiamonds in targeted intracellular labeling or biomolecule delivery.

doi: 10.1021/nn901014j

Acknowledgement
We are grateful to Jean-François Roch and Karen Perronet for fruitful discussions. We thank Géraldine Dantelle for the measurements of diamond colloidal suspensions zeta potentials and Anne Tarrade for the preparation of the samples observed with Transmission Electron Microscopy. This work was supported by the European Commission through the project Nano4Drugs" (contract LSHB-2005-CT-019102), by Agence Nationale de la Recherche through the project NaDia" (contract ANR-2007-PNANO-045) and by a Ile-de-France Region C'Nano grant under the project Biodiam".
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