Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Ultrafast ultrasound localization microscopy for deep super-resolution vascular imaging


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.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type



Prices may be subject to local taxes which are calculated during checkout

Figure 1: Principle of uULM.
Figure 2: Spatial resolution and quantification of uULM in the rat brain cortex through a thinned skull window.
Figure 3: uULM of the rat brain through a thinned skull window or through the intact skull.

Similar content being viewed by others


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

    Article  CAS  ADS  Google Scholar 

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

    Article  CAS  ADS  Google Scholar 

  3. Huang, B., Babcock, H. & Zhuang, X. Breaking the diffraction barrier: super-resolution imaging of cells. Cell 143, 1047–1058 (2010)

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  5. Couture, O., Tanter, M. & Fink, M. Method and device for ultrasound imaging. Patent Cooperation Treaty (PCT)/FR2011/052810 (2010)

  6. Desailly, Y., Couture, O., Fink, M. & Tanter, M. Sono-activated ultrasound localization microscopy. Appl. Phys. Lett. 103, 174107 (2013)

    Article  ADS  Google Scholar 

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

    Article  Google Scholar 

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

    Article  Google Scholar 

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

    Article  CAS  ADS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  12. Gessner, R. C., Frederick, C. B., Foster, F. S. & Dayton, P. A. Acoustic angiography: a new imaging modality for assessing microvasculature architecture. Int. J. Biomed. Imaging 2013, 936593 (2013)

    Article  Google Scholar 

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

  14. Tanter, M., Bercoff, J., Sandrin, L. & Fink, M. Ultrafast compound imaging for 2-D motion vector estimation: application to transient elastography. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 49, 1363–1374 (2002)

    Article  Google Scholar 

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

    Article  Google Scholar 

  16. Viessmann, O. M., Eckersley, R. J., Christensen-Jeffries, K., Tang, M. X. & Dunsby, C. Acoustic super-resolution with ultrasound and microbubbles. Phys. Med. Biol. 58, 6447–6458 (2013)

    Article  CAS  Google Scholar 

  17. Christensen-Jeffries, K., Browning, R. J., Tang, M.-X., Dunsby, C. & Eckersley, R. J. In vivo acoustic super-resolution and super-resolved velocity mapping using microbubbles. IEEE Trans. Med. Imaging 34, 433–440 (2015)

    Article  Google Scholar 

  18. Paxinos, G. & Watson, C. The Rat Brain in Stereotaxic Coordinates 6th edn (Academic, 2006)

  19. Szabo, T. L. in Diagnostic Ultrasound Imaging (ed. Szabo, T. L. ) 337–380 (Academic, 2004)

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

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  23. Goss, S. A., Frizzell, L. A. & Dunn, F. Ultrasonic absorption and attenuation in mammalian tissues. Ultrasound Med. Biol. 5, 181–186 (1979)

    Article  CAS  Google Scholar 

  24. Pernot, M., Montaldo, G., Tanter, M. & Fink, M. ‘Ultrasonic stars’ for time-reversal focusing using induced cavitation bubbles. Appl. Phys. Lett. 88, 034102 (2006)

    Article  ADS  Google Scholar 

  25. O’Reilly, M. A. & Hynynen, K. A super-resolution ultrasound method for brain vascular mapping. Med. Phys. 40, 110701 (2013)

    Article  Google Scholar 

  26. Walker, W. F. & Trahey, G. E. A fundamental limit on delay estimation using partially correlated speckle signals. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 42, 301–308 (1995)

    Article  Google Scholar 

  27. Osmanski, B.-F., Pezet, S., Ricobaraza, A., Lenkei, Z. & Tanter, M. Functional ultrasound imaging of intrinsic connectivity in the living rat brain with high spatiotemporal resolution. Nature Commun. 5, 5023 (2014)

    Article  CAS  ADS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  30. Errico, C., Osmanski, B.-F., Pezet, S., Couture, O., Lenkei, Z. & Tanter, M. Transcranial functional ultrasound imaging of the brain using microbubble-enhanced ultrasensitive Doppler. NeuroImage 124, 752–761 (2015)

    Article  Google Scholar 

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

    Article  Google Scholar 

  32. Duck, F. A. Physical Properties of Tissues: A Comprehensive Reference Book (Academic, 2013)

Download references


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

Authors and Affiliations


Corresponding author

Correspondence to Mickael Tanter.

Ethics declarations

Competing interests

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

Extended data figures and tables

Extended Data Figure 1 Schema of the temporal and spatial localization of unique sources.

a, Stack of B-mode images. The region of interest corresponds to a region of 2 mm × 1.1 mm within the cortex. b, Spatiotemporal filtering of the B-mode images shows the presence of decorrelating microbubbles in each frame (1–4). c, The four representative frames are separated by 44 ms (1–4). d, Computed two-dimensional PSF of the rescaled and filtered ultrafast acquisitions. These echoes are then interpolated and the Cartesian coordinates of their centre is obtained (1–4). The summit of each two-dimensional Gaussian profile identifies the centroid of each separable source.

Source data

Extended Data Figure 2 uULM coronal scan (anterior–posterior) of the entire rat brain through a thinned skull window.

ai, The ultrasound probe was driven by a micro-step motor to perform uULM on different imaging planes separated by 500 μm. We reconstructed the vascularization of the rat brain at the following coordinates: Bregma −0.5 mm (a), −1 mm (b), −1.5 mm (c), −2 mm (d), −2.5 mm (e), −3 mm (f), −3.5 mm (g), −4 mm (h), −4.5 mm (i).

Extended Data Figure 3 Anterior–posterior scan of in-plane velocity maps of the rat forebrain through a thinned skull window.

ai, Velocity maps for the different coronal planes presented in Extended Data Fig. 2.

Related audio

Supplementary information

Individual microbubble tracks within the cortex

Individual microbubble tracks within the cortex. (AVI 5333 kb)

Ultrafast ultrasound localization microscopy over multiple coronal plane of the cortex

Ultrafast ultrasound localization microscopy over multiple coronal plane of the cortex. (AVI 3137 kb)

PowerPoint slides

Source data

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Errico, C., Pierre, J., Pezet, S. et al. Ultrafast ultrasound localization microscopy for deep super-resolution vascular imaging. Nature 527, 499–502 (2015).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing