Self-illuminating quantum dots for highly sensitive in vivo real-time luminescent mapping of sentinel lymph nodes

Abstract

Qiang Wu, Maoquan ChuSchool of Life Science and Technology, Tongji University, Shanghai, People's Republic of ChinaAbstract: Quantum dots (QDs) show promise as novel nanomaterials for sentinel lymph node (SLN) mapping through their use in noninvasive in vivo fluorescence imaging, and they have provided remarkable results. However, in vivo fluorescence imaging has limitations mainly reflected in the strong autofluorescence and low deepness of tissue penetration associated with this technique. Here, we report on the first use of self-illuminating QDs for mouse axillary SLN mapping by bioluminescence resonance energy transfer, which was found to overcome these limitations. We used CdTe/CdS QDs synthesized in aqueous solution to conjugate a mutant of the bioluminescent protein, Renilla reniformis luciferase. The nanobioconjugates obtained had an average hydrodynamic diameter of 19 nm, and their luminescence catalyzed by the substrate (coelenterazine) could penetrate into at least 20 mm of hairless pigskin, which could be observed using an in vivo imaging system equipped with a 700 nm emission filter. Conversely, the fluorescence of the nanobioconjugates penetrated no more than 10 mm of pigskin and was observed with a strong background. When 80 µL of the nanobioconjugates (containing about 0.5 µmol/L of QDs) and 5 µL of coelenterazine (1 µg/µL) were intradermally injected into a mouse paw, the axillary SLN could be imaged in real time without external excitation, and little background interference was detected. Furthermore, the decayed luminescence of QD-Luc8 in SLNs could be recovered after being intradermally reinjected with the coelenterazine. Our data showed that using self-illuminating QDs, as opposed to fluorescence QDs, has greatly enhanced sensitivity in SLN mapping, and that the SLN could be identified synchronously by the luminescence and fluorescence of the self-illuminating QDs.Keywords: self-illuminating quantum dots, bioluminescence resonance energy transfer, sentinel lymph node, mapping, sensitivity