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Coherent anti-Stokes Raman scattering microscopy of single nanodiamonds


Nanoparticles have attracted enormous attention for biomedical applications as optical labels, drug-delivery vehicles and contrast agents in vivo. In the quest for superior photostability and biocompatibility, nanodiamonds are considered one of the best choices due to their unique structural, chemical, mechanical and optical properties. So far, mainly fluorescent nanodiamonds have been utilized for cell imaging. However, their use is limited by the efficiency and costs in reliably producing fluorescent defect centres with stable optical properties. Here, we show that single non-fluorescing nanodiamonds exhibit strong coherent anti-Stokes Raman scattering (CARS) at the sp3 vibrational resonance of diamond. Using correlative light and electron microscopy, the relationship between CARS signal strength and nanodiamond size is quantified. The calibrated CARS signal in turn enables the analysis of the number and size of nanodiamonds internalized in living cells in situ, which opens the exciting prospect of following complex cellular trafficking pathways quantitatively.

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Figure 1: Raman micro-spectroscopy of nanodiamonds.
Figure 2: Optical extinction of single nanodiamonds versus size.
Figure 3: CARS micro-spectroscopy of single nanodiamonds.
Figure 4: Quantitative DIC microscopy of single nanodiamonds.
Figure 5: CARS imaging of single nanodiamonds in fixed and living cells.


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The authors acknowledge the Cardiff University Large Research Equipment Fund for providing high-resolution TEM facilities, and thank G. Lalev and K. Cleal for their assistance. This work was funded by the UK BBSRC Research Council (grant nos BB/J021008/1 and BB/H006575/1). P.B. acknowledges the UK EPSRC Research Council for her Leadership fellowship award (grant no. EP/I005072/1).

Author information




P.B. and W.L. conceived and designed the experiments, interpreted the results and wrote the manuscript. I.P. performed the CARS experiments and analysed the data. L.P. performed the optical extinction cross-section measurements and analysed the data. G.Z. performed the quantitative DIC experiments and analysed the data. E.T. performed the SEM experiments and analysed the data. O.W. provided the bulk diamond and nanodiamond materials. P.W. performed the cell culture work. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Paola Borri.

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The authors declare no competing financial interests.

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Pope, I., Payne, L., Zoriniants, G. et al. Coherent anti-Stokes Raman scattering microscopy of single nanodiamonds. Nature Nanotech 9, 940–946 (2014).

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