Letter | Published:

Ultrafast ultrasound localization microscopy for deep super-resolution vascular imaging

Nature volume 527, pages 499502 (26 November 2015) | Download Citation


Non-invasive imaging deep into organs at microscopic scales remains an open quest in biomedical imaging. Although optical microscopy is still limited to surface imaging owing to optical wave diffusion and fast decorrelation in tissue, revolutionary approaches such as fluorescence photo-activated localization microscopy led to a striking increase in resolution by more than an order of magnitude in the last decade1. In contrast with optics, ultrasonic waves propagate deep into organs without losing their coherence and are much less affected by in vivo decorrelation processes. However, their resolution is impeded by the fundamental limits of diffraction, which impose a long-standing trade-off between resolution and penetration. This limits clinical and preclinical ultrasound imaging to a sub-millimetre scale. Here we demonstrate in vivo that ultrasound imaging at ultrafast frame rates (more than 500 frames per second) provides an analogue to optical localization microscopy by capturing the transient signal decorrelation of contrast agents—inert gas microbubbles. Ultrafast ultrasound localization microscopy allowed both non-invasive sub-wavelength structural imaging and haemodynamic quantification of rodent cerebral microvessels (less than ten micrometres in diameter) more than ten millimetres below the tissue surface, leading to transcranial whole-brain imaging within short acquisition times (tens of seconds). After intravenous injection, single echoes from individual microbubbles were detected through ultrafast imaging. Their localization, not limited by diffraction, was accumulated over 75,000 images, yielding 1,000,000 events per coronal plane and statistically independent pixels of ten micrometres in size. Precise temporal tracking of microbubble positions allowed us to extract accurately in-plane velocities of the blood flow with a large dynamic range (from one millimetre per second to several centimetres per second). These results pave the way for deep non-invasive microscopy in animals and humans using ultrasound. We anticipate that ultrafast ultrasound localization microscopy may become an invaluable tool for the fundamental understanding and diagnostics of various disease processes that modify the microvascular blood flow, such as cancer, stroke and arteriosclerosis.

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  1. 1.

    & Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19, 780–782 (1994)

  2. 2.

    et al. Imaging intracellular fluorescent proteins at nanometer resolution. Science 313, 1642–1645 (2006)

  3. 3.

    , & Breaking the diffraction barrier: super-resolution imaging of cells. Cell 143, 1047–1058 (2010)

  4. 4.

    & Ultrafast imaging in biomedical ultrasound. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 61, 102–119 (2014)

  5. 5.

    , & Method and device for ultrasound imaging. Patent Cooperation Treaty (PCT)/FR2011/052810 (2010)

  6. 6.

    , , & Sono-activated ultrasound localization microscopy. Appl. Phys. Lett. 103, 174107 (2013)

  7. 7.

    et al. Ultrafast imaging of ultrasound contrast agents. Ultrasound Med. Biol. 35, 1908–1916 (2009)

  8. 8.

    et al. Measurement of cerebral blood volume in mouse brain regions using micro-computed tomography. Neuroimage 47, 1312–1318 (2009)

  9. 9.

    et al. High-resolution structural and functional assessments of cerebral microvasculature using 3D Gas ΔR2*-mMRA. PLoS One 8, e78186 (2013)

  10. 10.

    et al. Multifunctional in vivo vascular imaging using near-infrared II fluorescence. Nature Med. 18, 1841–1846 (2012)

  11. 11.

    et al. High-speed label-free functional photoacoustic microscopy of mouse brain in action. Nature Methods 12, 407–410 (2015)

  12. 12.

    , , & Acoustic angiography: a new imaging modality for assessing microvasculature architecture. Int. J. Biomed. Imaging 2013, 936593 (2013)

  13. 13.

    et al. Spatiotemporal clutter filtering of ultrafast ultrasound data highly increases Doppler and fUltrasound sensitivity. IEEE Trans. Med. Imaging PP, 2271–2285 (2015)

  14. 14.

    , , & Ultrafast compound imaging for 2-D motion vector estimation: application to transient elastography. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 49, 1363–1374 (2002)

  15. 15.

    et al. Coherent plane wave compounding for very high frame rate ultrasonography of rapidly moving targets. IEEE Trans. Med. Imaging 32, 1265–1276 (2013)

  16. 16.

    , , , & Acoustic super-resolution with ultrasound and microbubbles. Phys. Med. Biol. 58, 6447–6458 (2013)

  17. 17.

    , , , & In vivo acoustic super-resolution and super-resolved velocity mapping using microbubbles. IEEE Trans. Med. Imaging 34, 433–440 (2015)

  18. 18.

    & The Rat Brain in Stereotaxic Coordinates 6th edn (Academic, 2006)

  19. 19.

    in Diagnostic Ultrasound Imaging (ed.) 337–380 (Academic, 2004)

  20. 20.

    et al. Imaging pericytes and capillary diameter in brain slices and isolated retinae. Nature Protocols 9, 323–336 (2014)

  21. 21.

    & Control of brain capillary blood flow. J. Cereb. Blood Flow Metab. 32, 1167–1176 (2012)

  22. 22.

    et al. Simultaneous measurement of RBC velocity, flux, hematocrit and shear rate in vascular networks. Nature Methods 7, 655–660 (2010)

  23. 23.

    , & Ultrasonic absorption and attenuation in mammalian tissues. Ultrasound Med. Biol. 5, 181–186 (1979)

  24. 24.

    , , & Ultrasonic stars’ for time-reversal focusing using induced cavitation bubbles. Appl. Phys. Lett. 88, 034102 (2006)

  25. 25.

    & A super-resolution ultrasound method for brain vascular mapping. Med. Phys. 40, 110701 (2013)

  26. 26.

    & A fundamental limit on delay estimation using partially correlated speckle signals. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 42, 301–308 (1995)

  27. 27.

    , , , & Functional ultrasound imaging of intrinsic connectivity in the living rat brain with high spatiotemporal resolution. Nature Commun. 5, 5023 (2014)

  28. 28.

    et al. An EFSUMB introduction into Dynamic Contrast-Enhanced Ultrasound (DCE-US) for quantification of tumor perfusion. Ultraschall Med. 33, 344–351 (2012)

  29. 29.

    et al. Ultrafast compound Doppler imaging: providing full blood flow characterization. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 58, 134–147 (2011)

  30. 30.

    , , , , & Transcranial functional ultrasound imaging of the brain using microbubble-enhanced ultrasensitive Doppler. NeuroImage 124, 752–761 (2015)

  31. 31.

    et al. Functional ultrasound imaging of the brain. Nature Methods 8, 662–664 (2011)

  32. 32.

    Physical Properties of Tissues: A Comprehensive Reference Book (Academic, 2013)

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This work was supported principally by the Agence Nationale de la Recherche (ANR), within the project ANR MUSLI. We thank the Fondation Pierre-Gilles de Gennes for funding C.E. The laboratory was also supported by LABEX WIFI (Laboratory of Excellence ANR-10-LABX-24) within the French Program “Investments for the Future” under reference ANR-10-IDEX-0001-02 PSL*.

Author information

Author notes

    • Olivier Couture
    •  & Mickael Tanter

    These authors contributed equally to this work.


  1. INSERM, Institut Langevin, 1 rue Jussieu, 75005 Paris, France

    • Claudia Errico
    • , Juliette Pierre
    • , Yann Desailly
    • , Olivier Couture
    •  & Mickael Tanter
  2. Institut Langevin, ESPCI-ParisTech, PSL Research University, 1 rue Jussieu, 75005 Paris, France

    • Claudia Errico
    • , Juliette Pierre
    • , Yann Desailly
    • , Olivier Couture
    •  & Mickael Tanter
  3. CNRS UMR 7587, 1 rue Jussieu, 75005 Paris, France

    • Claudia Errico
    • , Juliette Pierre
    • , Yann Desailly
    • , Olivier Couture
    •  & Mickael Tanter
  4. CNRS, UMR 8249, 10 rue Vauquelin, 75005 Paris, France

    • Sophie Pezet
    •  & Zsolt Lenkei
  5. Brain Plasticity Unit, ESPCI-ParisTech, PSL Research University, 10 rue Vauquelin, 75005 Paris, France

    • Sophie Pezet
    •  & Zsolt Lenkei


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Competing interests

M.T. is a co-founder, shareholder and scientific advisor of Supersonic Imagine. All other authors declare no competing financial interests.

Corresponding author

Correspondence to Mickael Tanter.

Extended data

Supplementary information


  1. 1.

    Individual microbubble tracks within the cortex

    Individual microbubble tracks within the cortex.

  2. 2.

    Ultrafast ultrasound localization microscopy over multiple coronal plane of the cortex

    Ultrafast ultrasound localization microscopy over multiple coronal plane of the cortex.

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