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Ultrasonic neuromodulation by brain stimulation with transcranial ultrasound


Brain stimulation methods are indispensable to the study of brain function. They have also proven effective for treating some neurological disorders. Historically used for medical imaging, ultrasound (US) has recently been shown to be capable of noninvasively stimulating brain activity. Here we provide a general protocol for the stimulation of intact mouse brain circuits using transcranial US, and, using a traditional mouse model of epilepsy, we describe how to use transcranial US to disrupt electrographic seizure activity. The advantages of US for brain stimulation are that it does not necessitate surgery or genetic alteration, but it confers spatial resolutions superior to other noninvasive methods such as transcranial magnetic stimulation. With a basic working knowledge of electrophysiology, and after an initial setup, ultrasonic neuromodulation (UNMOD) can be implemented in less than 1 h. Using the general protocol that we describe, UNMOD can be readily adapted to support a broad range of studies on brain circuit function and dysfunction.

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Figure 1: Basic ultrasonic brain stimulation rig and UNMOD waveform generation.
Figure 2: Preparation of electromyographic recordings to monitor US-evoked stimulation of intact motor cortex.
Figure 3: Electrophysiological recordings in response to brain stimulation with transcranial pulsed ultrasound.
Figure 4: Induction and disruption of electrographic seizure activity using UNMOD.


  1. 1

    Leighton, T.G. What is ultrasound? Prog. Biophys. Mol. Biol. 93, 3–83 (2007).

    Article  PubMed  Google Scholar 

  2. 2

    Harvey, E.N. The effect of high frequency sound waves on heart muscle and other irritable tissues. Am. J. Physiol. 91, 284–290 (1929).

    Article  Google Scholar 

  3. 3

    Fry, F.J., Ades, H.W. & Fry, W.J. Production of reversible changes in the central nervous system by ultrasound. Science 127, 83–84 (1958).

    CAS  Article  PubMed  Google Scholar 

  4. 4

    Tyler, W.J. et al. Remote excitation of neuronal circuits using low-intensity, low-frequency ultrasound. PLoS ONE 3, e3511 (2008).

    Article  PubMed  PubMed Central  Google Scholar 

  5. 5

    Tufail, Y. et al. Transcranial pulsed ultrasound stimulates intact brain circuits. Neuron 66, 681–694 (2010).

    CAS  Article  PubMed  Google Scholar 

  6. 6

    Manlapaz, J.S., Astroem, K.E., Ballantine, H.T. Jr. & Lele, P.P. Effects of ultrasonic radiation in experimental focal epilepsy in the cat. Exp. Neurol. 10, 345–356 (1964).

    CAS  Article  PubMed  Google Scholar 

  7. 7

    Cazzullo, C.L. & Guareschi, A. Experimental epilepsy produced by ultrasonics. I. Semeiological, electroencephalographic and anatomicopathological observations following application of low dosage of ultrasonics on guinea pig encephalon with intact theca. Riv. Patol. Nerv. Ment. 74, 545–572 (1953).

    CAS  PubMed  Google Scholar 

  8. 8

    Allegranza, A. Effect of anticonvulsant and neuroplegic drugs on experimental epilepsy induced with ultrasonics. Rev. Neurol. (Paris) 94, 395–399 (1956).

    CAS  Google Scholar 

  9. 9

    Min, B.K. et al. Focused ultrasound-mediated suppression of chemically-induced acute epileptic EEG activity. BMC Neurosci. 12, 23 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  10. 10

    O'Brien, W.D. Jr. Ultrasound-biophysics mechanisms. Prog. Biophys. Mol. Biol. 93, 212–255 (2007).

    Article  PubMed  Google Scholar 

  11. 11

    Dinno, M.A. et al. The significance of membrane changes in the safe and effective use of therapeutic and diagnostic ultrasound. Phys. Med. Biol. 34, 1543–1552 (1989).

    CAS  Article  PubMed  Google Scholar 

  12. 12

    ter Haar, G. Therapeutic applications of ultrasound. Prog. Biophys. Mol. Biol. 93, 111–129 (2007).

    Article  PubMed  Google Scholar 

  13. 13

    Dalecki, D. Mechanical bioeffects of ultrasound. Annu. Rev. Biomed. Eng. 6, 229–248 (2004).

    CAS  Article  PubMed  Google Scholar 

  14. 14

    Fritsch, G. & Hitzig, E. Über die elektrische Erregbarkeit des Grosshirns. Arch. Anat. Physiol. 37, 300–332 (1870).

    Google Scholar 

  15. 15

    Miesenbock, G. The optogenetic catechism. Science 326, 395–399 (2009).

    Article  CAS  PubMed  Google Scholar 

  16. 16

    Deisseroth, K. Optogenetics. Nat. Methods 8, 26–29 (2011).

    CAS  Article  PubMed  Google Scholar 

  17. 17

    Kringelbach, M.L., Jenkinson, N., Owen, S.L. & Aziz, T.Z. Translational principles of deep brain stimulation. Nat. Rev. Neurosci. 8, 623–635 (2007).

    CAS  Article  PubMed  Google Scholar 

  18. 18

    Schwalb, J.M. & Hamani, C. The history and future of deep brain stimulation. Neurotherapeutics 5, 3–13 (2008).

    Article  PubMed  PubMed Central  Google Scholar 

  19. 19

    Histed, M.H., Bonin, V. & Reid, R.C. Direct activation of sparse, distributed populations of cortical neurons by electrical microstimulation. Neuron 63, 508–522 (2009).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  20. 20

    Ranck, J.B. Jr. Which elements are excited in electrical stimulation of mammalian central nervous system: a review. Brain Res. 98, 417–440 (1975).

    Article  PubMed  Google Scholar 

  21. 21

    Stoney, S.D. Jr., Thompson, W.D. & Asanuma, H. Excitation of pyramidal tract cells by intracortical microstimulation: effective extent of stimulating current. J. Neurophysiol. 31, 659–669 (1968).

    Article  PubMed  Google Scholar 

  22. 22

    Tolias, A.S. et al. Mapping cortical activity elicited with electrical microstimulation using FMRI in the macaque. Neuron 48, 901–911 (2005).

    CAS  Article  PubMed  Google Scholar 

  23. 23

    Grill, W.M., Norman, S.E. & Bellamkonda, R.V. Implanted neural interfaces: biochallenges and engineered solutions. Annu. Rev. Biomed. Eng. 11, 1–24 (2009).

    CAS  Article  PubMed  Google Scholar 

  24. 24

    Payne, N.A. & Prudic, J. Electroconvulsive therapy: Part I. A perspective on the evolution and current practice of ECT. J. Psychiatr. Practice 15, 346–368 (2009).

    Article  Google Scholar 

  25. 25

    Wagner, T., Valero-Cabre, A. & Pascual-Leone, A. Noninvasive human brain stimulation. Annu. Rev. Biomed. Eng. 9, 527–565 (2007).

    CAS  Article  PubMed  Google Scholar 

  26. 26

    Barker, A.T. The history and basic principles of magnetic nerve stimulation. Electroencephalogr. Clin. Neurophysiol. Suppl. 51, 3–21 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. 27

    Fregni, F. & Pascual-Leone, A. Technology insight: noninvasive brain stimulation in neurology-perspectives on the therapeutic potential of rTMS and tDCS. Nat. Clin. Practice Neurol. 3, 383–393 (2007).

    Article  Google Scholar 

  28. 28

    Hallett, M. Transcranial magnetic stimulation: a primer. Neuron 55, 187–199 (2007).

    CAS  Article  PubMed  Google Scholar 

  29. 29

    Zhang, Y.P. et al. Use of magnetic stimulation to elicit motor evoked potentials, somatosensory evoked potentials, and H-reflexes in non-sedated rodents. J. Neurosci. Methods 165, 9–17 (2007).

    Article  PubMed  Google Scholar 

  30. 30

    Ji, R.R. et al. Repetitive transcranial magnetic stimulation activates specific regions in rat brain. Proc. Natl. Acad. Sci. USA 95, 15635–15640 (1998).

    CAS  Article  PubMed  Google Scholar 

  31. 31

    Cambiaghi, M. et al. Brain transcranial direct current stimulation modulates motor excitability in mice. Eur. J. Neurosci. 31, 704–709 (2010).

    Article  PubMed  Google Scholar 

  32. 32

    Zhang, S., Yin, L. & Fang, N. Focusing ultrasound with an acoustic metamaterial network. Phys. Rev. Lett, 102, 194301–194304 (2009).

    Article  CAS  Google Scholar 

  33. 33

    Li, J., Fok, L., Yin, X., Bartal, G. & Zhang, X. Experimental demonstration of an acoustic magnifying hyperlens. Nat. Mater. 8, 931–934 (2009).

    CAS  Article  PubMed  Google Scholar 

  34. 34

    Ayling, O.G., Harrison, T.C., Boyd, J.D., Goroshkov, A. & Murphy, T.H. Automated light-based mapping of motor cortex by photoactivation of channelrhodopsin-2 transgenic mice. Nat. Methods 6, 219–224 (2009).

    CAS  Article  PubMed  Google Scholar 

  35. 35

    Luft, A.R. et al. Transcranial magnetic stimulation in the rat. Exp. Brain Res. 140, 112–121 (2001).

    CAS  Article  PubMed  Google Scholar 

  36. 36

    Nielsen, J.B., Perez, M.A., Oudega, M., Enriquez-Denton, M. & Aimonetti, J.M. Evaluation of transcranial magnetic stimulation for investigating transmission in descending motor tracts in the rat. Eur. J.Neurosci. 25, 805–814 (2007).

    CAS  Article  PubMed  Google Scholar 

  37. 37

    Heimburg, T. Lipid ion channels. Biophys. Chem. 150, 2–22 (2010).

    CAS  Article  PubMed  Google Scholar 

  38. 38

    Heimburg, T. & Jackson, A.D. On soliton propagation in biomembranes and nerves. Proc. Natl. Acad. Sci. USA 102, 9790–9795 (2005).

    CAS  Article  PubMed  Google Scholar 

  39. 39

    Griesbauer, J., Wixforth, A. & Schneider, M.F. Wave propagation in lipid monolayers. Biophys. J. 97, 2710–2716 (2009).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  40. 40

    Yoo, S.S. et al. Focused ultrasound modulates region-specific brain activity. Neuroimage 56, 1267–1275 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  41. 41

    Ang, E.S. Jr., Gluncic, V., Duque, A., Schafer, M.E. & Rakic, P. Prenatal exposure to ultrasound waves impacts neuronal migration in mice. Proc. Natl. Acad. Sci. USA 103, 12903–12910 (2006).

    CAS  Article  PubMed  Google Scholar 

  42. 42

    Martin, E., Jeanmonod, D., Morel, A., Zadicario, E. & Werner, B. High intensity focused ultrasound for non-invaisve functional neurosurgery. Ann. Neurol. 66, 858–861 (2009).

    Article  PubMed  Google Scholar 

  43. 43

    Hynynen, K. & Clement, G. Clinical applications of focused ultrasound-the brain. Int. J. Hyperthermia 23, 193–202 (2007).

    CAS  Article  PubMed  Google Scholar 

  44. 44

    Altland, O.D., Dalecki, D., Suchkova, V.N. & Francis, C.W. Low-intensity ultrasound increases endothelial cell nitric oxide synthase activity and nitric oxide synthesis. J. Thromb. Haemost. 2, 637–643 (2004).

    CAS  Article  PubMed  Google Scholar 

  45. 45

    Claes, L. & Willie, B. The enhancement of bone regeneration by ultrasound. Prog. Biophys. Mol. Biol. 93, 384–398 (2007).

    Article  PubMed  Google Scholar 

  46. 46

    Ebisawa, K. et al. Ultrasound enhances transforming growth factor beta-mediated chondrocyte differentiation of human mesenchymal stem cells. Tissue Eng. 10, 921–929 (2004).

    CAS  Article  PubMed  Google Scholar 

  47. 47

    Hsu, H.C. et al. Ultrasound induces cyclooxygenase-2 expression through integrin, integrin-linked kinase, Akt, NF-kappaB and p300 pathway in human chondrocytes. Cell Signal. 19, 2317–2328 (2007).

    CAS  Article  PubMed  Google Scholar 

  48. 48

    Reher, P., Doan, N., Bradnock, B., Meghji, S. & Harris, M. Effect of ultrasound on the production of IL-8, basic FGF and VEGF. Cytokine 11, 416–423 (1999).

    CAS  Article  PubMed  Google Scholar 

  49. 49

    Sant'Anna, E.F., Leven, R.M., Virdi, A.S. & Sumner, D.R. Effect of low intensity pulsed ultrasound and BMP-2 on rat bone marrow stromal cell gene expression. J. Orthop. Res. 23, 646–652 (2005).

    CAS  Article  PubMed  Google Scholar 

  50. 50

    Sena, K., Leven, R.M., Mazhar, K., Sumner, D.R. & Virdi, A.S. Early gene response to low-intensity pulsed ultrasound in rat osteoblastic cells. Ultrasound Med. Biol. 31, 703–708 (2005).

    Article  PubMed  Google Scholar 

  51. 51

    Mihran, R.T., Barnes, F.S. & Wachtel, H. Transient modification of nerve excitability in vitro by single ultrasound pulses. Biomed. Sci. Instrum. 26, 235–246 (1990).

    CAS  PubMed  Google Scholar 

  52. 52

    White, P.J., Clement, G.T. & Hynynen, K. Longitudinal and shear mode ultrasound propagation in human skull bone. Ultrasound Med. Biol. 32, 1085–1096 (2006).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  53. 53

    White, P.J., Clement, G.T. & Hynynen, K. Local frequency dependence in transcranial ultrasound transmission. Phys. Med. Biol. 51, 2293–2305 (2006).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  54. 54

    Hayner, M. & Hynynen, K. Numerical analysis of ultrasonic transmission and absorption of oblique plane waves through the human skull. J. Acoust. Soc. Am. 110, 3319–3330 (2001).

    CAS  Article  PubMed  Google Scholar 

  55. 55

    Kinoshita, M., McDannold, N., Jolesz, F.A. & Hynynen, K. Targeted delivery of antibodies through the blood-brain barrier by MRI-guided focused ultrasound. Biochem. Biophys. Res. Commun. 340, 1085–1090 (2006).

    CAS  Article  PubMed  Google Scholar 

  56. 56

    Larrat, B. et al. MR-guided transcranial brain HIFU in small animal models. Phys. Med. Biol. 55, 365–388 (2010).

    CAS  Article  PubMed  Google Scholar 

  57. 57

    Hynynen, K., McDannold, N., Sheikov, N.A., Jolesz, F.A. & Vykhodtseva, N. Local and reversible blood-brain barrier disruption by noninvasive focused ultrasound at frequencies suitable for trans-skull sonications. Neuroimage 24, 12–20 (2005).

    Article  PubMed  Google Scholar 

  58. 58

    Hynynen, K. et al. Pre-clinical testing of a phased array ultrasound system for MRI-guided noninvasive surgery of the brain—a primate study. Eur. J. Radiol. 59, 149–156 (2006).

    Article  PubMed  Google Scholar 

  59. 59

    Hynynen, K. et al. 500-element ultrasound phased array system for noninvasive focal surgery of the brain: a preliminary rabbit study with ex vivo human skulls. Magn. Reson. Med. 52, 100–107 (2004).

    Article  PubMed  Google Scholar 

  60. 60

    Hynynen, K. & Jolesz, F.A. Demonstration of potential noninvasive ultrasound brain therapy through an intact skull. Ultrasound Med. Biol. 24, 275–283 (1998).

    CAS  Article  PubMed  Google Scholar 

  61. 61

    Clement, G.T. & Hynynen, K. A non-invasive method for focusing ultrasound through the human skull. Phys. Med. Biol. 47, 1219–1236 (2002).

    CAS  Article  PubMed  Google Scholar 

  62. 62

    Bartholow, R. Medical Electricity: a Practical Treatise on the Applications of Electricity to Medicine and Surgery 2nd edn, (Henry C. Lea's Son & Co., 1882).

  63. 63

    Barker, A.T., Jalinous, R. & Freeston, I.L. Non-invasive magnetic stimulation of human motor cortex. Lancet 1, 1106–1107 (1985).

    CAS  Article  PubMed  Google Scholar 

  64. 64

    Aravanis, A.M. et al. An optical neural interface: in vivo control of rodent motor cortex with integrated fiberoptic and optogenetic technology. J. Neural Eng. 4, S143–S156 (2007).

    Article  PubMed  Google Scholar 

  65. 65

    Penfield, W. & Jasper, H.H. Epilepsy and the Functional Anatomy of the Human Brain (J & A Churchill, 1954).

  66. 66

    Hamani, C., Andrade, D., Hodaie, M., Wennberg, R. & Lozano, A. Deep brain stimulation for the treatment of epilepsy. Int. J. Neural Syst. 19, 213–226 (2009).

    Article  PubMed  Google Scholar 

  67. 67

    Jobst, B.C. Electrical stimulation in epilepsy: vagus nerve and brain stimulation. Curr. Treat. Opt. Neurol. 12, 443–453 (2010).

    Article  Google Scholar 

  68. 68

    Theodore, W.H. & Fisher, R.S. Brain stimulation for epilepsy. Lancet Neurol. 3, 111–118 (2004).

    Article  PubMed  Google Scholar 

  69. 69

    Morrell, M. Brain stimulation for epilepsy: can scheduled or responsive neurostimulation stop seizures? Curr. Opin. Neurol. 19, 164–168 (2006).

    Article  PubMed  Google Scholar 

  70. 70

    Saillet, S. et al. Manipulating the epileptic brain using stimulation: a review of experimental and clinical studies. Epileptic Disord. 11, 100–112 (2009).

    PubMed  Google Scholar 

  71. 71

    Boon, P. et al. Deep brain stimulation in patients with refractory temporal lobe epilepsy. Epilepsia 48, 1551–1560 (2007).

    Article  PubMed  Google Scholar 

  72. 72

    Liebetanz, D. et al. Anticonvulsant effects of transcranial direct-current stimulation (tDCS) in the rat cortical ramp model of focal epilepsy. Epilepsia 47, 1216–1224 (2006).

    Article  PubMed  Google Scholar 

  73. 73

    Jennum, P. & Klitgaard, H. Repetitive transcranial magnetic stimulations of the rat. Effect of acute and chronic stimulations on pentylenetetrazole-induced clonic seizures. Epilepsy Res. 23, 115–122 (1996).

    CAS  Article  PubMed  Google Scholar 

  74. 74

    Sharma, A.K. et al. Mesial temporal lobe epilepsy: pathogenesis, induced rodent models and lesions. Toxicol. Pathol. 35, 984–999 (2007).

    Article  PubMed  Google Scholar 

  75. 75

    NEMA. Acoustic Output Measurement Standard For Diagnostic Ultrasound Equipment (National Electrical Manufacturers Association, 2004).

  76. 76

    Franklin, K.B.J. & Paxinos, G. The Mouse Brain in Stereotaxic Coordinates 3rd edn, (Academic Press, 2007).

  77. 77

    Racine, R.J. Modification of seizure activity by electrical stimulation. I. After-discharge threshold. Electroencephalogr. Clin. Neurophysiol. 32, 269–279 (1972).

    CAS  Article  PubMed  Google Scholar 

  78. 78

    Shealy, C.N. & Henneman, E. Reversible effects of ultrasound on spinal reflexes. Arch. Neurol. 374–386 (1962).

  79. 79

    Gavrilov, L.R. et al. The effect of focused ultrasound on the skin and deep nerve structures of man and animal. Prog. Brain Res. 43, 279–292 (1976).

    CAS  Article  PubMed  Google Scholar 

  80. 80

    Tsirulnikov, E.M. et al. Use of amplitude-modulated focused ultrasound for diagnosis of hearing disorders. Ultrasound Med. Biol. 14, 277–285 (1988).

    CAS  Article  PubMed  Google Scholar 

  81. 81

    Foster, K.R. & Wiederhold, M.L. Auditory responses in cats produced by pulsed ultrasound. J. Acoust. Soc. Am. 63, 1199–1205 (1978).

    CAS  Article  PubMed  Google Scholar 

  82. 82

    Magee, T.R. & Davies, A.H. Auditory phenomena during transcranial Doppler insonation of the basilar artery. J. Ultrasound Med. 12, 747–750 (1993).

    CAS  Article  PubMed  Google Scholar 

  83. 83

    Tsui, P.H., Wang, S.H. & Huang, C.C. In vitro effects of ultrasound with different energies on the conduction properties of neural tissue. Ultrasonics 43, 560–565 (2005).

    Article  PubMed  Google Scholar 

  84. 84

    Mihran, R.T., Barnes, F.S. & Wachtel, H. Temporally-specific modification of myelinated axon excitability in vitro following a single ultrasound pulse. Ultrasound Med. Biol. 16, 297–309 (1990).

    CAS  Article  PubMed  Google Scholar 

  85. 85

    Foley, J.L., Little, J.W. & Vaezy, S. Image-guided high-intensity focused ultrasound for conduction block of peripheral nerves. Ann. Biomed. Eng. 35, 109–119 (2007).

    Article  PubMed  Google Scholar 

  86. 86

    Bachtold, M.R., Rinaldi, P.C., Jones, J.P., Reines, F. & Price, L.R. Focused ultrasound modifications of neural circuit activity in a mammalian brain. Ultrasound Med. Biol. 24, 557–565 (1998).

    CAS  Article  PubMed  Google Scholar 

  87. 87

    Rinaldi, P.C., Jones, J.P., Reines, F. & Price, L.R. Modification by focused ultrasound pulses of electrically evoked responses from an in vitro hippocampal preparation. Brain Res. 558, 36–42 (1991).

    CAS  Article  PubMed  Google Scholar 

  88. 88

    Ludwig, G.D. The velocity of sound through tissues and the acoustic impedance of tissues. J. Acoust. Soc. Am. 22, 862–866 (1950).

    Article  Google Scholar 

  89. 89

    Turker, K.S. Electromyography: some methodological problems and issues. Phys. Ther. 73, 698–710 (1993).

    CAS  Article  PubMed  Google Scholar 

  90. 90

    Whelan, P.J. Electromyogram recordings from freely moving animals. Methods 30, 127–141 (2003).

    CAS  Article  PubMed  Google Scholar 

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Support for this work was provided by start-up funds from Arizona State University to W.J.T. and Department of Defense grants from the US Army Research, Development, and Engineering Command (RDECOM W911NF-09-0431) and a Defense Advanced Research Projects Agency Young Faculty Award (DARPA N66001-10-1-4032) to W.J.T.

Author information




Y.T., A.Y., S.P. and W.J.T. designed and conducted the experimental procedures. Y.T., A.Y., M.M.L. and W.J.T. analyzed and interpreted data from the experiments. Y.T., A.Y., S.P., M.M.L. and W.J.T. wrote the manuscript.

Corresponding author

Correspondence to William J Tyler.

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Competing interests

W.J.T. is a cofounder of SynSonix.

Supplementary information

Supplementary Video 1

Overview of ultrasonic neuromodulation rig and equipment (MOV 30900 kb)

Supplementary Video 2

Disruption of seizure activity with TPU (MOV 8962 kb)

Supplementary Video 3

Disruption of electrographic seizure activity with continuous wave UNMOD waveforms (MOV 1490 kb)

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Tufail, Y., Yoshihiro, A., Pati, S. et al. Ultrasonic neuromodulation by brain stimulation with transcranial ultrasound. Nat Protoc 6, 1453–1470 (2011).

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