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Self-illuminating quantum dot conjugates for in vivo imaging

Nature Biotechnology volume 24, pages 339–343 (2006)Cite this article

Abstract

Fluorescent semiconductor quantum dots hold great potential for molecular imaging in vivo1,2,3,4,5. However, the utility of existing quantum dots for in vivo imaging is limited because they require excitation from external illumination sources to fluoresce, which results in a strong autofluorescence background and a paucity of excitation light at nonsuperficial locations. Here we present quantum dot conjugates that luminesce by bioluminescence resonance energy transfer in the absence of external excitation. The conjugates are prepared by coupling carboxylate-presenting quantum dots to a mutant of the bioluminescent protein Renilla reniformis luciferase. We show that the conjugates emit long-wavelength (from red to near-infrared) bioluminescent light in cells and in animals, even in deep tissues, and are suitable for multiplexed in vivo imaging. Compared with existing quantum dots, self-illuminating quantum dot conjugates have greatly enhanced sensitivity in small animal imaging, with an in vivo signal-to-background ratio of > 103 for 5 pmol of conjugate.

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Figure 1: Design and spectroscopic characterization of bioluminescent quantum dot conjugates based on BRET.
Figure 2: Bioluminescence and fluorescence imaging of QD655-Luc8 and Luc8 injected subcutaneously (I and II) and intramuscularly (III and IV) at indicated sites in a mouse (I and III, QD655-Luc8, 5 pmol; II and IV, Luc8, 30 pmol).
Figure 3: Multiplexed imaging of conjugates QD605-Luc8, QD655-Luc8, QD705-Luc8 and QD800-Luc8 in vitro and in mice.
Figure 4: Imaging C6 glioma cells labeled with QD655-Luc8-R9 in vitro and in mice.

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Acknowledgements

This work was supported by the Burroughs Wellcome Fund (to J.R.), the Korea Research Foundation Grant M07-2004-000-10234-0 (to M.-K.S.), a Stanford Bio-X Graduate Fellowship (to A.M.L.), the National Institutes of Health grants 5R01CA82214-7 and P50 CA114747 (to S.S.G.), the National Cancer Institute Centers of Cancer Nanotechnology Excellence (CCNE) U54, and the National Cancer Institute's Small Animal Imaging Resource Program (SAIRP R24CA92862).

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Author notes

  1. Min-Kyung So and Chenjie Xu: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Radiology & Bio-X Program, Molecular Imaging Program at Stanford, Stanford University, 1201 Welch Road, Stanford, 94305-5484, California, USA

    Min-Kyung So, Chenjie Xu, Andreas M Loening, Sanjiv S Gambhir & Jianghong Rao

  2. Department of Bioengineering, Stanford University, 1201 Welch Road, Stanford, 94305-5484, California, USA

    Andreas M Loening & Sanjiv S Gambhir

  3. Biophysics Program, Stanford University, 1201 Welch Road, Stanford, 94305-5484, California, USA

    Jianghong Rao

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  1. Min-Kyung So
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Correspondence to Jianghong Rao.

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Supplementary information

Supplementary Fig. 1

Dependence of the BRET efficiency on the distance between QDs and Luc8's. (PDF 60 kb)

Supplementary Fig. 2

Effect of the number of immobilized Luc8 on the BRET efficiency. (PDF 89 kb)

Supplementary Fig. 3

Bioluminescence and fluorescence imaging of QD655-Luc8 and Luc8 in a living mouse (BRET filter: 650–660 nm). (PDF 45 kb)

Supplementary Table 1

The BRET ratio of QD-Luc8 conjugates. (PDF 12 kb)

Supplementary Notes (PDF 73 kb)

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So, MK., Xu, C., Loening, A. et al. Self-illuminating quantum dot conjugates for in vivo imaging. Nat Biotechnol 24, 339–343 (2006). https://doi.org/10.1038/nbt1188

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