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Diamond nanoparticles (nanodiamonds) can transport active drugs in culturedcells as well as in vivo. However, in the latter case, methods allowing todetermine their bioavailability accurately are still lacking. Nanodiamond canbe made fluorescent with a perfectly stable emission and a lifetime ten timeslonger than the one of tissue autofluorescence. Taking advantage of theseproperties, we present an automated quantification method of fluorescentnanodiamonds (FND) in histological sections of mouse organs and tumor, aftersystemic injection. We use a home-made time-delayed fluorescence microscopecomprising a custom pulsed laser source synchronized on the master clock of agated intensified array detector. This setup allows to obtainultra-high-resolution images 120 Mpixels of whole mouse organs sections, withsubcellular resolution and single-particle sensitivity. As a proof-of-principleexperiment, we quantified the biodistribution and aggregation state of newcationic FNDs able to transport small interfering RNA inhibiting the oncogeneresponsible for Ewing sarcoma. Image analysis showed a low yield ofnanodiamonds in the tumor after intravenous injection. Thus, for the in vivoefficacy assay we injected the nanomedicine into the tumor. We achieved a28-fold inhibition of the oncogene. This method can readily be applied to othernanoemitters with $\approx$100 ns lifetime.

nanoscale

Harnessing subcellular-resolved organ distribution of cationic  copolymer-functionalized fluorescent nanodiamonds for optimal delivery of  therapeutic siRNA to a xenografted tumor in mice