← Explore

Optoacoustic brain stimulation at submillimeter spatial precision

Ying Jiang, Hyeon Jeong Lee, Lu Lan, Hua-an Tseng, Chen Yang, Heng-Ye Man, Xue Han, Ji-Xin Cheng

Nature Communications 2020, 11 · 10.1038/s41467-020-14706-1

rodentin vitro cellhealthycellular imaginginvasive electrophysiologyemg mephistology molecular

Abstract

Low-intensity ultrasound is an emerging modality for neuromodulation. Yet, transcranial neuromodulation using low-frequency piezo-based transducers offers poor spatial confinement of excitation volume, often bigger than a few millimeters in diameter. In addition, the bulky size limits their implementation in a wearable setting and prevents integration with other experimental modalities. Here, we report spatially confined optoacoustic neural stimulation through a miniaturized Fiber-Optoacoustic Converter (FOC). The FOC has a diameter of 600 μm and generates omnidirectional ultrasound wave locally at the fiber tip through the optoacoustic effect. We show that the acoustic wave generated by FOC can directly activate individual cultured neurons and generate intracellular Ca 2+ transients. The FOC activates neurons within a radius of 500 μm around the fiber tip, delivering superior spatial resolution over conventional piezo-based low-frequency transducers. Finally, we demonstrate direct and spatially confined neural stimulation of mouse brain and modulation of motor activity in vivo.

Abstract via europepmc.

Speciesmouse (C57BL/6J, in vivo); rat (primary cortical neuron culture, in vitro)
Subjects3, 3, 2swept animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlinactive transducer, active control site, other
Auditory controlcontrol experiment
Readout timingonline
Anaesthesiaanaesthetised
Readoutscellular imaging, invasive electrophysiology, emg mep, histology molecularcalcium imaging (OGD-1) in cultured neurons/glia; local field potential (LFP) in S1/A1; EMG from forelimb triceps brachii; post-hoc H&E histology
Direction of effectexcitatoryFOC (fiber-optoacoustic converter) stimulation produced calcium transients in cultured neurons, LFP responses in S1, and EMG responses (motor activation) in forelimb muscle, consistent with direct neural activation.
Adverse eventsnone observedNo obvious morphological changes were detected in neurons stimulated multiple times over a 2-h duration; no tissue damage was observed on histology after repeated in vivo stimulation.

Exposures

Exposure 1: FOC stimulation of cultured cortical neurons (in vitro)

Target: cultured neurons — “cultured rat cortical neurons
Device: custom-built · Fiber-Optoacoustic Converter (FOC)

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500, 5,000swept?
Pulse duration (ms)0.001✓✓
Pulse repetition frequency (Hz)3,600✓✓
Duty cycle (%)0.36pulse duration × PRF gives 0.36%✓✓
Sonication duration (s)0.02, 0.05, 0.1, 0.2swept✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)480✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatenot applicable
In-situ pressure (kPa)not applicable
In-situ Isppa (W/cm²)not applicable
In-situ Ispta (W/cm²)not applicable
Protocol, in the paper’s words

The FOC emits omnidirectional 1-us acoustic pulses at a 3.6 kHz laser repetition rate (duty cycle ~0.36%). Stimulation trains of 20, 50, 100 or 200 ms were delivered to cultured neurons to find an activation threshold (100 and 50 ms produced calcium transients, 20 ms did not); repeated reliability was also tested with eight 200-ms bursts at a 2-s inter-burst interval.

Exposure 2: FOC stimulation of mouse primary somatosensory cortex (S1), in vivo

Target: primary somatosensory cortex — “primary somatosensory cortex (S1)
Device: custom-built · Fiber-Optoacoustic Converter (FOC)

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500, 5,000swept?
Pulse duration (ms)0.001✓✓
Pulse repetition frequency (Hz)3,600✓✓
Duty cycle (%)0.36pulse duration × PRF gives 0.36%✓✓
Sonication duration (s)0.05, 0.2swept✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)480✓✓
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
Protocol, in the paper’s words

FOC placed in contact with the exposed brain surface (craniotomy, dura intact, saline-immersed); 200 ms and 50 ms laser pulse trains delivered while LFP was recorded from S1 with a tungsten electrode; separate trials tested ipsilateral A1 as an anatomical control and delivery without brain contact as a negative control.

Exposure 3: FOC stimulation of mouse primary motor cortex (M1), in vivo

Target: primary motor cortex — “motor cortex
Device: custom-built · Fiber-Optoacoustic Converter (FOC)

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500, 5,000swept?
Pulse duration (ms)0.001✓✓
Pulse repetition frequency (Hz)3,600✓✓
Duty cycle (%)0.36pulse duration × PRF gives 0.36%✓✓
Sonication duration (s)0.2✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)480✓✓
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
Protocol, in the paper’s words

A 200 ms laser pulse train was delivered to the FOC placed on the motor cortex (stereotaxic coordinates) while EMG was recorded from the contralateral forelimb triceps brachii; the FOC was scanned over a 3x3 mm^2 grid (1 mm spacing) to map motor representation.

Flags from extraction

  • n_subjectsPaper never states a single total number of mice used across all in vivo experiments; group sizes stated per sub-experiment are 3 mice (S1/A1 LFP recordings), 3 mice (M1 EMG mapping), 2 mice (S1-to-forelimb EMG latency); these may partly overlap and are listed as reported rather than summed.
  • exposures[0].fundamental_frequency_khzThe FOC is a broadband optoacoustic source (no discrete carrier/fundamental frequency); the stated 0.5-5 MHz range from the RF spectrum is recorded as the frequency range rather than a single value.
  • exposures[0].free_field.pressure_kpa0.48 MPa is a general device characterization (measured underwater with a needle hydrophone at the fiber tip) rather than a value measured for each specific in vitro/in vivo experiment; applied to all three exposures as the only reported pressure figure.
  • sham_typeControls used were mechanistic (FOC lifted off contact / no acoustic coupling; uncoated fiber with laser only; FOC aimed at a different site (A1) for spatial-confinement testing) rather than a classic transducer-off sham; mapped to inactive_transducer/active_control_site/other.
  • auditory_controlPaper tests for (and rules out) an indirect auditory/cochlear pathway by recording contralateral A1 responses, but does not implement a masking-sound or similar auditory control procedure; mapped to 'other'.