  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
