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Monitoring cerebral hemodynamic change during transcranial ultrasound stimulation using optical intrinsic signal imaging

Evgenii Kim, Eloise Anguluan, Jae Gwan Kim

Scientific Reports 2017, 7 · 10.1038/s41598-017-13572-0

rodenthealthycerebral haemodynamics

Abstract

Transcranial ultrasound stimulation (tUS) is a promising non-invasive approach to modulate brain circuits. The application is gaining popularity, however the full effect of ultrasound stimulation is still unclear and further investigation is needed. This study aims to apply optical intrinsic signal imaging (OISI) for the first time, to simultaneously monitor the wide-field cerebral hemodynamic change during tUS on awake animal with high spatial and temporal resolution. Three stimulation paradigms were delivered using a single-element focused transducer operating at 425 kHz in pulsed mode having the same intensity (I SPPA = 1.84 W/cm 2 , I SPTA = 129 mW/cm 2 ) but varying pulse repetition frequencies (PRF). The results indicate a concurrent hemodynamic change occurring with all actual tUS but not under a sham stimulation. The stimulation initiated the increase of oxygenated hemoglobin (HbO) and decrease of deoxygenated hemoglobin (RHb). A statistically significant difference (p < 0.05) was found in the amplitude change of hemodynamics evoked by varying PRF. Moreover, the acoustic stimulation was able to trigger a global as well as local cerebral hemodynamic alteration in the mouse cortex. Thus, the implementation of OISI offers the possibility of directly investigating brain response in an awake animal during tUS through cerebral hemodynamic change.

Abstract via europepmc.

Speciesmouse (BALB/c)
Subjects12 animals
Sessions per subject1
Randomisedyes
Blindingnot reported
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingboth
Anaesthesiaboth
Readoutscerebral haemodynamicsoptical intrinsic signal imaging (OISI) of cortical HbO/RHb hemodynamics; laser speckle imaging (LSI) of cerebral blood flow (separate sub-experiment, n=3 mice)
Direction of effectexcitatoryAll three tUS paradigms (differing only in PRF/pulse duration at matched Isppa/Ispta) increased HbO and decreased RHb relative to sham, indicating activation; peak HbO amplitude was significantly larger with higher PRF (shorter pulses).
Adverse eventsnot reportedThe sonication parameters were selected to be below FDA diagnostic-ultrasound limits (Ispta 720 mW/cm2, Isppa 190 W/cm2, MI 1.9) and the peak negative pressure of 0.53 MPa used was described as far below the threshold for thermal or cavitation-related brain tissue damage.

Exposures

Exposure 1: tUS of mouse cortex at three PRF/pulse-duration paradigms (matched Isppa/Ispta)

Target: cerebral cortex — “mouse cortex, directed towards the bregma to provide a full image of the cortical brain
Device: Olympus / Panametrics · Olympus Corp. · V301-SU

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)425✓✓
Pulse duration (ms)0.188, 0.094, 0.047swept✓✓
Pulse repetition frequency (Hz)375, 750, 1,500swept✓✓
Duty cycle (%)not reported
Sonication duration (s)0.2✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)530✓✓
Free-field Isppa (W/cm²)1.84✓✓
Free-field Ispta (W/cm²)0.129✓✓
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

The ultrasound delivered was a 425 kHz pulse-modulated signal containing 10 pulse trains given over a period of 5 seconds for each trial. Every stimulus is a pulse train lasting 200 ms with M pulses given at a rate determined by the pulse repetition frequency (PRF), with each pulse having N cycles (Table 1: PRF 375/750/1500 Hz, N cycles 80/40/20, M pulses 75/150/300). Each trial image acquisition lasts for a total of 17 s including 5 s of stimulation containing ten ultrasound pulse trains with a repetition rate of 2 Hz; the interval between the beginning of each trial was fixed to 1 minute. The experiment on awake animals consisted of 100 randomly shuffled trials (25 per paradigm, including sham).

Consistency checks: intensity pressure inconsistent free field.

Flags from extraction

  • exposures[0].timing.pulse_duration_msPulse duration not stated directly in ms; computed from N cycles ÷ fundamental frequency (Table 1: N=80/40/20 cycles at 425 kHz), per the sanctioned cycles-per-pulse conversion.
  • exposures[0].unspecified_domainThe paper does not state whether the reported Isppa/Ispta/pressure values were measured in water (free field) or in situ; classified as unspecified domain.
  • exposures[0].timingThe protocol nests short pulses (intra-train PRF 375/750/1500 Hz) within 200 ms trains that repeat 10 times over 5 s at a 2 Hz train-repetition rate; the schema's single PRF/duty-cycle fields cannot represent both levels. pulse_duration_ms and pulse_repetition_frequency_hz refer to the intra-train pulsing; sonication_duration_s is set to the 200 ms train length per the paper's own definition of a pulse train.
  • randomisedRandomisation described is of trial order/paradigm sequence within a single-arm, within-subject design (100 randomly shuffled trials), not allocation to separate treatment groups.