Technical Report | Published:

Multifunctional in vivo vascular imaging using near-infrared II fluorescence

Nature Medicine volume 18, pages 18411846 (2012) | Download Citation

Abstract

In vivo real-time epifluorescence imaging of mouse hind limb vasculatures in the second near-infrared region (NIR-II) is performed using single-walled carbon nanotubes as fluorophores. Both high spatial (30 μm) and temporal (<200 ms per frame) resolution for small-vessel imaging are achieved at 1–3 mm deep in the hind limb owing to the beneficial NIR-II optical window that affords deep anatomical penetration and low scattering. This spatial resolution is unattainable by traditional NIR imaging (NIR-I) or microscopic computed tomography, and the temporal resolution far exceeds scanning microscopic imaging techniques. Arterial and venous vessels are unambiguously differentiated using a dynamic contrast-enhanced NIR-II imaging technique on the basis of their distinct hemodynamics. Further, the deep tissue penetration and high spatial and temporal resolution of NIR-II imaging allow for precise quantifications of blood velocity in both normal and ischemic femoral arteries, which are beyond the capabilities of ultrasonography at lower blood velocities.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    et al. Carotid-artery intima and media thickness as a risk factor for myocardial infarction and stroke in older adults. N. Engl. J. Med. 340, 14–22 (1999).

  2. 2.

    , , & Carotid artery wall thickness: comparison between sonography and multi-detector row CT angiography. Neuroradiology 52, 75–82 (2010).

  3. 3.

    , , , & Magnetic resonance images lipid, fibrous, calcified, hemorrhagic, and thrombotic components of human atherosclerosis in vivo. Circulation 94, 932–938 (1996).

  4. 4.

    et al. Ultrasound biomicroscopy in small animal research: applications in molecular and preclinical imaging. J. Biomed. Biotechnol. 2012 10.1155/2012/519238 (2012).

  5. 5.

    & 3-dimensional reconstruction from projections—review of algorithms. Int. Rev. Cytol. 38, 111–151 (1974).

  6. 6.

    et al. Visualization of advanced human prostate cancer lesions in living mice by a targeted gene transfer vector and optical imaging. Nat. Med. 8, 891–897 (2002).

  7. 7.

    et al. A route to brightly fluorescent carbon nanotubes for near-infrared imaging in mice. Nat. Nanotechnol. 4, 773–780 (2009).

  8. 8.

    , & Deep-tissue anatomical imaging of mice using carbon nanotube fluorophores in the second near-infrared window. Proc. Natl. Acad. Sci. USA 108, 8943–8948 (2011).

  9. 9.

    et al. In vivo biodistribution and highly efficient tumour targeting of carbon nanotubes in mice. Nat. Nanotechnol. 2, 47–52 (2007).

  10. 10.

    et al. High performance in vivo near-IR (>1 μm) imaging and photothermal cancer therapy with carbon nanotubes. Nano Res. 3, 779–793 (2010).

  11. 11.

    , , & Carbon nanotubes in biology and medicine: in vitro and in vivo detection, imaging and drug delivery. Nano Res. 2, 85–120 (2009).

  12. 12.

    , , , & Carbon materials for drug delivery & cancer therapy. Mater. Today 14, 316–323 (2011).

  13. 13.

    et al. Near-infrared fluorescent type II quantum dots for sentinel lymph node mapping. Nat. Biotechnol. 22, 93–97 (2004).

  14. 14.

    , , & In vivo imaging of tumors with protease-activated near-infrared fluorescent probes. Nat. Biotechnol. 17, 375–378 (1999).

  15. 15.

    et al. Receptor-targeted optical imaging of tumors with near-infrared fluorescent ligands. Nat. Biotechnol. 19, 327–331 (2001).

  16. 16.

    et al. In vivo near-infrared fluorescence imaging of osteoblastic activity. Nat. Biotechnol. 19, 1148–1154 (2001).

  17. 17.

    et al. Near-infrared-fluorescence-enhanced molecular imaging of live cells on gold substrates. Angew. Chem. Int. Ed. Engl. 50, 4644–4648 (2011).

  18. 18.

    et al. Three-dimensional imaging of single nanotube molecule endocytosis on plasmonic substrates. Nat. Comm. 3, 700 (2012).

  19. 19.

    , & Bioimaging: Second window for in vivo imaging. Nat. Nanotechnol. 4, 710–711 (2009).

  20. 20.

    et al. Laser-induced tissue hyperthermia mediated by gold nanoparticles: toward cancer phototherapy. J. Biomed. Opt. 14, 021016 (2009).

  21. 21.

    et al. Selection of quantum dot wavelengths for biomedical assays and imaging. Mol. Imaging 2, 50–64 (2003).

  22. 22.

    In vivo near-infrared fluorescence imaging. Curr. Opin. Chem. Biol. 7, 626–634 (2003).

  23. 23.

    & All-optical anatomical co-registration for molecular imaging of small animals using dynamic contrast. Nat. Photonics 1, 526–530 (2007).

  24. 24.

    et al. A pilot toxicology study of single-walled carbon nanotubes in a small sample of mice. Nat. Nanotechnol. 3, 216–221 (2008).

  25. 25.

    et al. Circulation and long-term fate of functionalized, biocompatible single-walled carbon nanotubes in mice probed by Raman spectroscopy. Proc. Natl. Acad. Sci. USA 105, 1410–1415 (2008).

  26. 26.

    , , & The largest and smallest X-ray computed-tomography systems. Nucl. Instrum. Meth. A 221, 207–212 (1984).

  27. 27.

    Current status of developments and applications of micro-CT. Annu. Rev. Biomed. Eng. 13, 531–552 (2011).

  28. 28.

    et al. Embryonic stem cell–derived endothelial cells engraft into the ischemic hindlimb and restore perfusion. Arterioscl. Throm. Vasc. Biol. 30, 984–991 (2010).

  29. 29.

    , & Simulations of axial mixing of liquids in a long horizontal pipe for industrial applications. Energy Fuels 24, 5844–5850 (2010).

  30. 30.

    et al. Endothelial cells derived from human iPSCS increase capillary density and improve perfusion in a mouse model of peripheral arterial disease. Arterioscl. Throm. Vasc. Biol. 31, e72–e79 (2011).

  31. 31.

    et al. Assessment of mouse hind limb endothelial function by measuring femoral artery blood flow responses. J. Vasc. Surg. 53, 1350–1358 (2011).

  32. 32.

    & Iodinated contrast media and their adverse reactions. J. Nucl. Med. Technol. 36, 69–74 (2008).

  33. 33.

    et al. Evaluation of the radiation dose in micro-CT with optimization of the scan protocol. Contrast Media Mol. Imaging 5, 201–207 (2010).

  34. 34.

    et al. Near-infrared photoluminescent Ag2S quantum dots from a single source precursor. J. Am. Chem. Soc. 132, 1470–1471 (2010).

  35. 35.

    et al. Ag2S quantum dot: a bright and biocompatible fluorescent nanoprobe in the second near-infrared window. ACS Nano 6, 3695–3702 (2012).

  36. 36.

    , , & Optical investigation of infrared dyes in sol–gel films. J. Lumin. 87–89, 748–750 (2000).

  37. 37.

    , , & Optical properties of single-walled carbon nanotubes separated in a density gradient: length, bundling, and aromatic stacking effects. J. Phys. Chem. C Nanomater. Interfaces 114, 19569–19575 (2010).

  38. 38.

    , , & Preparation of carbon nanotube bioconjugates for biomedical applications. Nat. Protoc. 4, 1372–1382 (2009).

  39. 39.

    , , & Murine model of hindlimb ischemia. J. Vis. Exp. 23, 1035 (2009).

  40. 40.

    International Commission on Non-Ionizing Radiation Protection. Revision of guidelines on limits of exposure to laser radiation of wavelengths between 400 nm and 1.4 μm. Health Phys. 79, 431–440 (2000).

Download references

Acknowledgements

This study was supported by grants from the National Cancer Institute of the US National Institutes of Health to H.D. (5R01CA135109-02), the National Heart, Lung and Blood Institute of the US National Institutes of Health to J.P.C. (U01HL100397, RC2HL103400) and N.F.H. (K99HL098688) and a Stanford Graduate Fellowship to G.H.

Author information

Author notes

    • Guosong Hong
    •  & Jerry C Lee

    These authors contributed equally to this work.

Affiliations

  1. Department of Chemistry, Stanford University, Stanford, California, USA.

    • Guosong Hong
    • , Joshua T Robinson
    • , Liming Xie
    •  & Hongjie Dai
  2. Division of Cardiovascular Medicine, School of Medicine, Stanford University, Stanford, California, USA.

    • Jerry C Lee
    • , Uwe Raaz
    • , Ngan F Huang
    •  & John P Cooke

Authors

  1. Search for Guosong Hong in:

  2. Search for Jerry C Lee in:

  3. Search for Joshua T Robinson in:

  4. Search for Uwe Raaz in:

  5. Search for Liming Xie in:

  6. Search for Ngan F Huang in:

  7. Search for John P Cooke in:

  8. Search for Hongjie Dai in:

Contributions

H.D., J.P.C., N.F.H., G.H. and J.C.L. conceived of and designed the experiments. G.H., J.C.L., J.T.R., U.R., L.X. and N.F.H. performed the experiments. G.H., J.C.L., U.R., L.X., N.F.H., J.P.C. and H.D. analyzed the data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Ngan F Huang or John P Cooke or Hongjie Dai.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6, Supplementary Tables 1,2

Videos

  1. 1.

    Supplementary Movie 1

    Carbon nanotube labeled blood flow in a healthy limb. NIR-II video-rate imaging showing blood flow in a healthy hind limb of a control mouse. Total movie time: 37.5 s, frame rate: 5.3 frames/s.

  2. 2.

    Supplementary Movie 2

    Carbon nanotube labeled blood flow in an ischemic limb early frames. NIR-II video-rate imaging showing significantly reduced blood flow in an ischemic hind limb with occlusion in the femoral artery. The same number of frames as in Supplementary Movie 1 is shown in this movie to highlight the dramatic delay of flow into femoral artery and vein. Total movie time: 37.5 s, frame rate: 5.3 frames/s.

  3. 3.

    Supplementary Movie 3

    Carbon nanotube labeled blood flow in an ischemic limb late frames. NIR-II video-rate imaging showing significantly reduced blood flow in an ischemic hind limb with occlusion in the femoral artery up to 247.5 s post injection of carbon nanotubes. Total movie time: 247.5 s, frame rate: 0.89 frames/s.

  4. 4.

    Supplementary Movie 4

    Differentiation of arterial and venous vessels subserving a larger area. NIR-II video-rate imaging showing different blood flow behaviors of arterial and venous vessels subserving a larger area than just the hind limb, which is the basis for differentiation of blood vessel type based on principal component analysis. Total movie time: 31.9 s, frame rate: 5.3 frames/s.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nm.2995

Further reading