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Miniature ultrasound ring array transducers for transcranial ultrasound neuromodulation of freely-moving small animals

Hyunggug Kim, Seongyeon Kim, Nam Suk Sim, Cristina Pasquinelli, Axel Thielscher, Jeong Ho Lee, Hyunjoo J. Lee

Brain Stimulation 2019, 12, 251-255 · 10.1016/j.brs.2018.11.007

rodenthealthyemg mepbehaviour

Abstract

Background Current transcranial ultrasound stimulation for small animal in vivo experiment is limited to acute stimulation under anesthesia in stereotaxic fixation due to bulky and heavy curved transducers. Methods We developed a miniaturized ultrasound ring array transducer which is capable of invoking motor responses through neuromodulation of freely-moving awake mice. Results The developed transducer is a 32-element, 183-kHz ring array with a weight of 0.035 g (with PCB: 0.73 g), a diameter of 8.1 mm, a focal length of 2.3 mm, and lateral resolution of 2.75 mm. By developing an affixation scheme suitable for freely-moving animals, the transducer was successfully coupled to the mouse brain and induced motor responses in both affixed and awake states. Conclusion Ultrasound neuromodulation of a freely-moving animal is now possible using the developed lightweight and compact system to conduct a versatile set of in vivo experiments.

Abstract via europepmc.

Speciesmouse
Subjects4 animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlinactive transducer
Auditory controlsound only sham
Readout timingonline
Anaesthesianot reported
Readoutsemg mep, behaviourEMG recording of motor responses (success defined as 3x EMG noise floor); visual/video observation of motor responses in freely-moving mice
Direction of effectexcitatoryTranscranial ultrasound to motor cortex via a miniature CMUT ring array evoked EMG motor responses with success rate increasing with intensity (Ispta), reaching over 70% average success at 34.1 mW/cm2 in 4 mice; success rate was negligible or much reduced with no AC drive, no DC bias, or a flipped (aimed-away) transducer.
Adverse eventsnone observedNo significant tissue damage such as vascular hemorrhage or neuronal necrosis was observed compared to control mice.

Exposures

Exposure 1: Transcranial ultrasound stimulation of motor cortex (acute and freely-moving mice)

Target: motor cortex — “motor cortex
Device: custom-built · 32-element, 183-kHz CMUT ring array transducer

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)183✓✓
Pulse duration (ms)4.5✓✓
Pulse repetition frequency (Hz)200✓✓
Duty cycle (%)90pulse duration × PRF gives 90%✓✓
Sonication duration (s)0.2✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Ispta, domain unspecified (W/cm²)0.0341, 0.0554swept?
Protocol, in the paper’s words

The CMUT array was biased at DC bias voltage of 100 V and driven with AC peak-to-peak voltage of 80 V at 183 kHz. Each stimulation trial consisted of 40-pulses of 90% duty cycle at a pulse repeat frequency (PRF) of 200 Hz, and each 4.5-ms long pulse consisted of 756 pulses of ultrasound. At each intensity, approximately 25 stimulation trials were conducted over 4 min, with success rate measured by adjusting the AC voltage to vary intensity. Since the duration of sonication in the in vivo protocol is only 0.2 s per trial, the temperature effect (~0.1C after ~240 s of continuous sonication in a phantom) should be negligible.

Flags from extraction

  • n_subjectsText explicitly gives 'four mice' for the main intensity-response experiment, but additional single-mouse control experiments (0V AC, 0V DC, flipped device) and the separate freely-moving-mouse demonstration may or may not overlap with these four animals; total unique subjects across all experiments is unclear.
  • anaesthesiaThe paper contrasts prior work using anesthetized, head-fixed animals with its own acute and awake freely-moving experiments, but never explicitly states whether the acute (head-fixed) mouse experiments were performed under anesthesia.
  • exposures[0].unspecified_domain.ispta_w_cm2Intensity values (55.4 mW/cm2 at the described 80V driving condition; 34.1 mW/cm2 at which success rate exceeded 70%) are not stated as free-field or in-situ; domain is unspecified. Figure 2 caption labels the varied intensity as ISPPA while the text calls the same quantity ISPTA, a text/figure inconsistency.
  • sham_typeThree control conditions are described (0V AC with 100V DC; 0V DC with 80V AC; device flipped/aimed away); classified overall as inactive_transducer, with the flipped-device condition also serving as an auditory (buzzing sound) sham.