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Frequency Dependence of Ultrasound Neurostimulation in the Mouse Brain

Patrick Peiyong Ye, Julian R. Brown, Kim Butts Pauly

Ultrasound in Medicine & Biology 2016, 42, 1512-1530 · 10.1016/j.ultrasmedbio.2016.02.012

rodenthealthyemg mep

Abstract

Ultrasound neuromodulation holds promise as a non-invasive technique for neuromodulation of the central nervous system. However, much remains to be determined about how the technique can be transformed into a useful technology, including the effect of ultrasound frequency. Previous studies have demonstrated neuromodulation in vivo using frequencies <1 MHz, with a trend toward improved efficacy with lower frequency. However, using higher frequencies could offer improved ultrasound spatial resolution. We investigate the ultrasound neuromodulation effects in mice at various frequencies both below and above 1 MHz. We find that frequencies up to 2.9 MHz can still be effective for generating motor responses, but we also confirm that as frequency increases, sonications require significantly more intensity to achieve equivalent efficacy. We argue that our results provide evidence that favors either a particle displacement or a cavitation-based mechanism for the phenomenon of ultrasound neuromodulation.

Abstract via europepmc.

Speciesmouse (C57BL/6)
Subjects5, 5, 5, 5, 6swept animals
Sessions per subjectnot reported
Randomisedyes
Blindingnot reported
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingonline
Anaesthesiaanaesthetised
Readoutsemg mepForelimb (triceps) EMG: success rate of evoked contractions, contraction latency, contraction duration, peak amplitude
Direction of effectexcitatoryUltrasound from 0.3 to 2.9 MHz evoked forelimb muscle contractions (measured by EMG); success rate and efficacy decreased and required intensity increased substantially as frequency increased, with success rate correlating with particle displacement and cavitation index but not radiation force.
Adverse eventsnot reported

Exposures

Exposure 1: Frequency sweep (0.3-2.9 MHz) ultrasound stimulation of mouse motor cortex

Target: motor cortex — “motor cortex (forelimb representation)
Device: Olympus / Panametrics · Olympus

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)300, 400, 500, 600, 1,000, 1,400, 1,900, 2,400, 2,900swept✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)0.08✓✓
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 estimatederatingmean or range across subjects
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)0.02, 127swept✓✓
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Five sets of experiments (A-E) tested continuous-wave sonications at frequencies from 0.3 to 2.9 MHz. Experiment A compared 0.3-0.6 MHz using both a constant 40,000-cycle duration and a constant 80 ms duration; experiments B-E used a fixed 80 ms CW pulse duration throughout (0.6-1.4 MHz in B, 1.4-2.9 MHz in C, 0.3-0.6 MHz with a different focused transducer in D, and 1.4 vs 2.9 MHz mapped across multiple brain sites in E). Within each block, sonications of different intensities plus one sham (transducer output off) were delivered in random order, with at least a 2 s delay between sonications.

Flags from extraction

  • n_subjectsGroup sizes are given per sub-experiment (A-D: n=5 mice each, E: n=6 mice); the paper never states a combined total, and some mice/experiments were later excluded (see Methods), so group sizes are listed rather than summed.
  • exposures[0].target.termsTarget is the mouse motor cortex region controlling forelimb (triceps) contraction, identified via EMG; not a more specific sub-region such as primary motor cortex.
  • exposures[0].fundamental_frequency_khzFrequencies pooled from Table 1 across five sub-experiments (A-E) that used different transducers/geometries; treated as one exposure per the frequency-response-curve exception since target and general timing (80 ms CW pulses) were held constant across the sweep.
  • exposures[0].device.modelFour different Olympus transducers (0.5 MHz planar 1 in.; 1 MHz focused 0.75 in., 1.90 in. PTF; 2.25 MHz focused 0.75 in., 1 in. PTF used for both experiments C and E) were used across the frequency sweep; see paper Table 1 for exact geometry per experiment.
  • exposures[0].in_situ.isppa_w_cm2Value is an approximate overall range ('about 1' to 'more than 100 W/cm2') stated in the Results text for threshold intensities across the full 0.3-2.9 MHz sweep; per-frequency/per-experiment ranges differ substantially and are given only in Table 1 and Figs. 5a-d, which could not be captured by a single verbatim quote.
  • exposures[0].in_situ.isppa_w_cm2Domain classified as in_situ because intensities are described in figure captions (e.g., Fig. 5) as 'spatial peak intensity after accounting for skull attenuation', i.e. derated for measured mouse-skull insertion loss.