Stimulating Electrodes Research Articles

The Induction of Long-Term Potentiation by Medial Septum Activation under Urethane Anesthesia Can Alter Gene Expression in the Hippocampus.

We studied changes in the expression of early genes in hippocampal cells in response to stimulation of the dorsal medial septal area (dMSA), leading to long-term potentiation in the hippocampus. Rats under urethane anesthesia were implanted with stimulating electrodes in the ventral hippocampal commissure and dMSA and a recording electrode in the CA1 area of the hippocampus. We found that high-frequency stimulation (HFS) of the dMSA led to the induction of long-term potentiation in the synapses formed by the ventral hippocampal commissure on the hippocampal CA1 neurons. One hour after dMSA HFS, we collected the dorsal and ventral hippocampi on both the ipsilateral (damaged by the implanted electrode) and contralateral (intact) sides and analyzed the expression of genes by qPCR. The dMSA HFS led to an increase in the expression of bdnf and cyr61 in the ipsilateral hippocampi and egr1 in the ventral contralateral hippocampus. Thus, dMSA HFS under the conditions of degeneration of the cholinergic neurons in the medial septal area prevented the described increase in gene expression. The changes in cyr61 expression appeared to be dependent on the muscarinic M1 receptors. Our data suggest that the induction of long-term potentiation by dMSA activation enhances the expression of select early genes in the hippocampus.

Recurrent laryngeal motor nerve conduction studies in a rat model: Establishing an objective measure for investigating laryngeal innervation.

Disease or injury can cause neuromuscular changes to the larynx that can affect voice, breathing, and swallowing. Motor nerve conduction studies have had limited use in the study of laryngeal neurophysiology, despite their importance in other anatomic sites. The aim of this study was to explore the feasibility of performing recurrent laryngeal motor nerve conduction studies (rlMNCS) in a rat model. rlMNCS were performed in 15 rats under anesthesia. A bipolar stimulating electrode was placed on the recurrent laryngeal nerve (RLN) 5 mm below the cricoid cartilage. Via direct laryngoscopy, a recording electrode was placed transorally into the thyroarytenoid muscle. The RLN was maximally stimulated to determine the compound muscle action potential (CMAP). Three consecutive trials were averaged. The mean stimulating threshold to the RLN to achieve a CMAP from the thyroarytenoid was 1.7 ± 0.6 mA. RLN stimulation caused a visible adductor twitch of the vocal fold in all animals. The mean negative amplitude was 2.0 ± 0.8 mV, and the total area was 1.0 ± 0.4 mV ms. The CMAP latency and negative duration were 1.0 ± 0.1 ms and 0.9 ± 0.2 ms, respectively. rlMNCS are feasible and may be useful in understanding laryngeal neurophysiology with disease or injury. This work could provide a tractable animal model for studying and monitoring treatment of neuromuscular conditions affecting voice, breathing, and swallowing.

A self-powered biocompatible brain probe for remote blood pressure regulation

Hypertension, a common cardiovascular disorder characterized by elevated blood pressure levels, is conventionally managed using pharmacological interventions to achieve optimal blood pressure control. In this study, a novel self-powered, biocompatible brain probe is presented as a potential long-term and remote approach to blood pressure regulation. The device consists of a piezoelectric transducer, an electronic module, brain stimulating electrodes, and a drug microneedle array. As the resonator, the piezoelectric transducer is embedded in resonant structure within resin shell, enabling the harvesting of acoustic energy from smartphone audio tone without any external power sources. By programming specific audio signals, the resonator can be activated, and the electronic module can output corresponding electrostimulation signals on demand. Implanted brain stimulating electrodes in the ventrolateral periaqueductal grey matter (vlPAG) of rats can be wirelessly controlled by the smartphone, resulting in a significant reduction in blood pressure of approximately 20 mmHg. The device's drug microneedle array, located beneath the pedestal of device, can deliver antibiotics percutaneously to prevent post-surgical infections, indicating a biocompatible approach for long-term hypertension treatment. This novel, self-powered and wireless treatment has significant potential for clinical hypertension therapy and may broaden the scope of self-powered techniques for telemedicine and brain-machine interfaces.

Impedance spectroscopy of enlarged cochlear implant stimulation electrodes – FEM simulations considering the perilymph

Abstract Cochlear implants are hearing prostheses for patients with severe to total hearing loss but intact auditory nerve. An external speech processor captures sound from the environment, which is subsequently converted into electrical signals and transmitted to an electrode array in the patient’s inner ear. The metallic stimulation electrodes of the electrode array electrically stimulate the spiral ganglion cells of the auditory nerve. The functionality of cochlear implants strongly depends on the possible maximum current stimulating the spiral ganglion cells, which can be affected by, e.g., cell growth around the stimulation electrodes. This in turn decreases the stimulation efficiency leading to decreased hearing. Cell growth, implant position and other changes in the surrounding medium are reflected in a change of the impedance of the stimulation electrodes. The impedance measurement of the stimulation electrodes is already implemented in all common cochlear implant systems to check functionality of the stimulation electrodes after implantation, but the frequency spectrum is normally not analyzed. Although this method can detect cell growth on the stimulation electrodes, it faces limitations when other interfering effects, such as changes in the perilymph and implant position, influence the impedance. This work shows impedance spectroscopic measurements using enlarged cochlear implant models to electrically analyze the surrounding medium, the perilymph, to understand changes in electrode impedance and to later monitor the stimulation efficiency of cochlear implants and to identify possible reasons for decreased hearing ability by impedance spectroscopy. In addition, we use FEM simulations to numerically model the influence of the perilymph composition on the impedance measurement. As shown by a final validation, this model can serve as a basis for an extended simulation model including implant position and cell growth monitoring to predict hearing deterioration in cochlear implant patients. In this context, this work serves as a basis for the development of a holistic prediction model and considers in the first step exclusively the influence of the perilymph composition on the impedance between two stimulation electrodes.

Modulating individual axons and axonal populations in the peripheral nerve using transverse intrafascicular multichannel electrodes.

Objective.A transverse intrafascicular multichannel electrode (TIME) may offer advantages over more conventional cuff electrodes including higher spatial selectivity and reduced stimulation charge requirements. However, the performance of TIME, especially in the context of non-conventional stimulation waveforms, remains relatively unexplored. As part of our overarching goal of investigating stimulation efficacy of TIME, we developed a computational toolkit that automates the creation and usage ofin siliconerve models with TIME setup, which solves nerve responses using cable equations and computes extracellular potentials using finite element method.Approach.We began by implementing a flexible and scalable Python/MATLAB-based toolkit for automatically creating models of nerve stimulation in the hybrid NEURON/COMSOL ecosystems. We then developed a sciatic nerve model containing 14 fascicles with 1,170 myelinated (A-type, 30%) and unmyelinated (C-type, 70%) fibers to study fiber responses over a variety of TIME arrangements (monopolar and hexapolar) and stimulation waveforms (kilohertz stimulation and cathodic ramp modulation).Main results.Our toolkit obviates the conventional need to re-create the same nerve in two disparate modeling environments and automates bi-directional transfer of results. Our population-based simulations suggested that kilohertz stimuli provide selective activation of targeted C fibers near the stimulating electrodes but also tended to activate non-targeted A fibers further away. However, C fiber selectivity can be enhanced by hexapolar TIME arrangements that confined the spatial extent of electrical stimuli. Improved upon prior findings, we devised a high-frequency waveform that incorporates cathodic DC ramp to completely remove undesirable onset responses.Conclusion.Our toolkit allows agile, iterative design cycles involving the nerve and TIME, while minimizing the potential operator errors during complex simulation. The nerve model created by our toolkit allowed us to study and optimize the design of next-generation intrafascicular implants for improved spatial and fiber-type selectivity.

Postoperative Respiratory Dysfunction in Patients with Parkinson’s Disease

This case report presents a 67-year-old female with Parkinson’s disease who underwent deep brain stimulation (DBS) electrode replacement surgery. Following extubation, the patient developed persistent laryngospasm, requiring immediate intervention. Suspecting symptoms similar to levodopa withdrawal, the patient received intravenous propofol and enteral levodopa supplementation, leading to symptom improvement. The case underscores the importance of perioperative management, including timely medication supplementation and DBS functioning, in Parkinson’s patients to prevent neuroleptic malignant syndrome (NMS)-like complications. Anesthesiologists should be vigilant about potential airway issues and NMS in this population and maintain optimal fluid status during surgery. Prompt intervention can prevent irreversible damage and improve patient outcomes.

Vagus Nerve Stimulation Electrode Impedance Over Time in Children With Lennox-Gastaut Syndrome

ObjectiveThis manuscript describes the behavior of impedance of vagus nerve stimulation (VNS) electrode over time in a cohort of children with Lennox-Gastaut syndrome. Materials and MethodsNineteen consecutive pediatric patients with Lennox-Gastaut syndrome submitted to VNS were studied. All patients had at least four years of follow-up. Serial impedance measurements were carried out during every out-patient visit. A baseline value was obtained one month after surgery, before generator activation and yearly values were recorded for the next four years. Outcome regarding seizures was obtained through analysis of standardized seizure diaries filled out by the patient, relatives, or caregivers. ResultsThere were 12 boys. Age ranged from four to 14 years (mean = 7.2). Mean impedance value was 2635 Ω at baseline, 2576 Ω after one year, 2418 Ω after two years, 2340 Ω after three years, and 2241 Ω after four years. There was a mean impedance decrease of 17% after four years. This decrease was statistically significant compared with baseline by the second year of follow-up: p = 0.342 after one year, p = 0.007 after two years, p = 0.001 after three years, and p = 0.001 after four years. There was no significant relationship between impedance values and seizure outcome at any time point. ConclusionsVNS electrode impedance significantly decreased during long-term follow-up in children with Lennox-Gastaut syndrome. To our knowledge, this is the first report on such findings regarding VNS in the literature. These findings suggest that the electrode/nerve interface is stable during long-term follow-up of VNS therapy and that this preserved anatomical relationship might be related to our ability to safely stimulate and review/explant the system whenever needed.

Electrochemical Properties of Sputtered Ruthenium Oxide Neural Stimulation and Recording Electrodes

A chronically stable electrode material with a low impedance for recording neural activity, and a high charge-injection capacity for functional electro-stimulation is desirable for the fabrication of implantable microelectrode arrays that aim to restore impaired or lost neurological functions in humans. For this purpose, we have investigated the electrochemical properties of sputtered ruthenium oxide (RuOx) electrode coatings deposited on planar microelectrode arrays, using an inorganic model of interstitial fluid (model-ISF) at 37 °C as the electrolyte. Through a combination of cyclic voltammetry (CV) and an electrochemical impedance spectroscopy (EIS) modelling study, we have established the contribution of the faradaic reaction as the major charge-injection contributor within the safe neural stimulation potential window of ±0.6 V vs. Ag|AgCl. We have also established the reversibility of the charge-injection process for sputtered RuOx film, by applying constant charge-per-phase current stimulations at different pulse widths, and by comparing the magnitudes of the leading and trailing access voltages during voltage transient measurements. Finally, the impedance of the sputtered RuOx film was found to be reasonably comparable in both its oxidized and reduced states, although the electronic contribution from the capacitive double-layer was found to be slightly higher for the completely oxidized film around 0.6 V than for its reduced counterpart around −0.6 V.

Evaluation of the PaCER Algorithm for Postoperative Subthalamic Nucleus Deep Brain Stimulation Electrode Localization.

Deep Brain Stimulation (DBS) is an established therapy for many movement disorders. DBS entails electrical stimulation of precise brain structures using permanently implanted electrodes. Following implantation, locating the electrodes relative to the target brain structure assists patient outcome optimization. Here we evaluated an open-source automatic algorithm (PaCER) to localize individual electrodes on Computed Tomography imaging (co-registered to Magnetic Resonance Imaging). In a dataset of 111 participants, we found a modified version of the algorithm matched manual-markups with median error less than 0.191 mm (interquartile range 0.698 mm). Given the error is less than the voxel resolution (1 mm3) of the images, we conclude that the automatic algorithm is suitable for DBS electrode localizations.Clinical Relevance- Automated DBS electrode localization identifies the closest electrode to the target brain structure; allowing the neurologist to direct electrical stimulation to maximize patient outcomes. Further, if none of the electrodes are deemed suitable, localization will guide re-implantation.

Vagus nerve stimulation using an endovascular electrode array

Objective. Vagus nerve stimulation (VNS), which involves a surgical procedure to place electrodes directly on the vagus nerve (VN), is approved clinically for the treatment of epilepsy, depression, and to facilitate rehabilitation in stroke. VNS at surgically implanted electrodes is often limited by activation of motor nerve fibers near and within the VN that cause neck muscle contraction. In this study we investigated endovascular VNS that may allow activation of the VN at locations where the motor nerve fibers are not localized. Approach. We used endovascular electrodes within the nearby internal jugular vein (IJV) to electrically stimulate the VN while recording VN compound action potentials (CAPs) and neck muscle motor evoked potentials (MEPs) in an acute intraoperative swine experiment. Main Results. We show that the stimulation electrode position within the IJV is critical for efficient activation of the VN. We also demonstrate use of fluoroscopy (cone beam CT mode) and ultrasound to determine the position of the endovascular stimulation electrode with respect to the VN and IJV. At the most effective endovascular stimulation locations tested, thresholds for VN activation were several times higher than direct stimulation of the nerve using a cuff electrode; however, this work demonstrates the feasibility of VNS with endovascular electrodes and provides tools to optimize endovascular electrode positions for VNS. Significance. This work lays the foundation to develop endovascular VNS strategies to stimulate at VN locations that would be otherwise too invasive and at VN locations where structures such as motor nerve fibers do not exist.

Velocity of conduction between columns and layers in barrel cortex reported by parvalbumin interneurons.

Inhibitory interneurons expressing parvalbumin (PV) play critical roles throughout the brain. Their rapid spiking enables them to control circuit dynamics on a millisecond time scale, and the timing of their activation by different excitatory pathways is critical to these functions. We used a genetically encoded hybrid voltage sensor to image PV interneuron voltage changes with sub-millisecond precision in primary somatosensory barrel cortex (BC) of adult mice. Electrical stimulation evoked depolarizations with a latency that increased with distance from the stimulating electrode, allowing us to determine conduction velocity. Spread of responses between cortical layers yielded an interlaminar conduction velocity and spread within layers yielded intralaminar conduction velocities in different layers. Velocities ranged from 74 to 473μm/ms depending on trajectory; interlaminar conduction was 71% faster than intralaminar conduction. Thus, computations within columns are more rapid than between columns. The BC integrates thalamic and intracortical input for functions such as texture discrimination and sensory tuning. Timing differences between intra- and interlaminar PV interneuron activation could impact these functions. Imaging of voltage in PV interneurons reveals differences in signaling dynamics within cortical circuitry. This approach offers a unique opportunity to investigate conduction in populations of axons based on their targeting specificity.

How far does electrical stimulation activate white matter tracts? A computational modeling study

ObjectiveThe aim of this study was to model how the different parameters of electrical stimulation (intensity, pulse shape, probe geometry) influence the extent of white matter activation. MethodsThe electrical potentials generated by the stimulating electrodes were determined by solving Laplace equation. The temporal evolution of membrane potentials at each nodes of Ranvier of an axon was then computed by solving the coupled system of differential equations describing membrane dynamics and cable propagation. ResultsRegions of unilateral propagation were observed for monophasic pulses delivered with a bipolar probe aligned along the tract. For biphasic pulses, the largest activation areas and depths were found with a high inter-electrode-distance (IED) bipolar probe, oriented orthogonally to the tract. The smallest activation areas and depths were found for bipolar stimulations with the probe aligned parallel to the tract and low IED. For isotropic white matter regions, the activation area and depth were three times larger than for anisotropic white matter tracts. ConclusionsBipolar probes with biphasic pulses offer the greatest versatility: an orthogonal orientation acts as two monopolars (increased sensitivity when searching for a tract), whereas a parallel orientation corresponds to a single monopolar (increased specificity). Activation is more superficial when stimulating highly anisotropic tracts. SignificanceThis knowledge is essential for interpreting the behavorial effects of stimulation and the recordings of axono-cortical evoked potentials.

Delayed closed-loop neurostimulation for the treatment of pathological brain rhythms in mental disorders: a computational study.

Mental disorders are among the top most demanding challenges in world-wide health. A large number of mental disorders exhibit pathological rhythms, which serve as the disorders characteristic biomarkers. These rhythms are the targets for neurostimulation techniques. Open-loop neurostimulation employs stimulation protocols, which are rather independent of the patients health and brain state in the moment of treatment. Most alternative closed-loop stimulation protocols consider real-time brain activity observations but appear as adaptive open-loop protocols, where e.g., pre-defined stimulation sets in if observations fulfil pre-defined criteria. The present theoretical work proposes a fully-adaptive closed-loop neurostimulation setup, that tunes the brain activities power spectral density (PSD) according to a user-defined PSD. The utilized brain model is non-parametric and estimated from the observations via magnitude fitting in a pre-stimulus setup phase. Moreover, the algorithm takes into account possible conduction delays in the feedback connection between observation and stimulation electrode. All involved features are illustrated on pathological α- and γ-rhythms known from psychosis. To this end, we simulate numerically a linear neural population brain model and a non-linear cortico-thalamic feedback loop model recently derived to explain brain activity in psychosis.

Computer vision-based algorithm to sUppoRt coRrect electrode placemeNT (CURRENT) for home-based electric non-invasive brain stimulation

ObjectiveHome-based non-invasive brain stimulation (NIBS) has been suggested as an adjunct treatment strategy for neuro-psychiatric disorders. There are currently no available solutions to direct and monitor correct placement of the stimulation electrodes. To address this issue, we propose an easy-to-use digital tool to support patients for self-application. MethodsWe recruited 36 healthy participants and compared their cap placement performance with the one of a NIBS-expert investigator. We tested participants’ placement accuracy with instructions before (Pre) and after the investigator’s placement (Post), as well as participants using the support tool (CURRENT). User experience (UX) and confidence were further evaluated. ResultsPermutation tests demonstrated a smaller deviation within the CURRENT compared with Pre cap placement (p = 0.02). Subjective evaluation of ease of use and usefulness of the tool were vastly positive (8.04 out of 10). CURRENT decreased the variability of performance, ensured placement within the suggested maximum of deviation (10 mm) and supported confidence of correct placement. ConclusionsThis study supports the usability of this novel technology for correct electrode placement during self-application in home-based settings. SignificanceCURRENT provides an exciting opportunity to promote home-based, self-applied NIBS as a safe, high-frequency treatment strategy that can be well integrated in patients’ daily lives.

Practice of anaesthesia for electroconvulsive therapy

Electroconvulsive therapy (ECT) is an established therapeutic method for the treatment of severe mental disorders refractory to pharmaco- and psychotherapy. ECT is a first-line treatment option in delusional disorders, severe depression with acute suicidal tendency or life-threatening catatonia. Usually, ECT is performed as a treatment series. Under short-term anaesthesia and muscle relaxation, tonic-clonic seizures are induced using an external stimulation electrode. Convulsion can be exerted by uni- or bipolar stimulation using an electric charge up to 1000millicoulomb (mC) with an amperage of 900mA. Muscular relaxation is necessary to prevent injuries caused by uncontrolled movements during convulsion. During paralysis, consciousness is blocked by general anaesthesia, although ECT is associated with antegrade amnesia for seizure induction and the seizure itself. In the context of ECT, the ideal hypnotic should be characterised by rapid onset, short duration of action and negligible anticonvulsive effects (i.e., least possible impact on seizure quality and duration). As mutual awareness of psychiatric and anaesthesiologic techniques is essential for safe and effective conduction of ECT, this article presents ECT both from the psychiatrist's and the anaesthesiologist's perspective.

Quantifying the Variability Associated with Postoperative Localization of Deep Brain Stimulation Electrodes

Introduction: Computational models of deep brain stimulation (DBS) have become common tools in clinical research studies that attempt to establish correlations between stimulation locations in the brain and behavioral outcome measures. However, the accuracy of any patient-specific DBS model depends heavily upon accurate localization of the DBS electrodes within the anatomy, which is typically defined via co-registration of clinical CT and MRI datasets. Several different approaches exist for this challenging registration problem, and each approach will result in a slightly different electrode localization. The goal of this study was to better understand how different processing steps (e.g., cost-function masking, brain extraction, intensity remapping) affect the estimate of the DBS electrode location in the brain. Methods: No “gold standard” exists for this kind of analysis, as the exact location of the electrode in the living human brain cannot be determined with existing clinical imaging approaches. However, we can estimate the uncertainty associated with the electrode position, which can be used to guide statistical analyses in DBS mapping studies. Therefore, we used high-quality clinical datasets from 10 subthalamic DBS subjects and co-registered their long-term postoperative CT with their preoperative surgical targeting MRI using 9 different approaches. The distances separating all of the electrode location estimates were calculated for each subject. Results: On average, electrodes were located within a median distance of 0.57 mm (0.49–0.74) of one another across the different registration approaches. However, when considering electrode location estimates from short-term postoperative CTs, the median distance increased to 2.01 mm (1.55–2.78). Conclusions: The results of this study suggest that electrode location uncertainty needs to be factored into statistical analyses that attempt to define correlations between stimulation locations and clinical outcomes.

Fabrication of a transparent array of penetrating 3D microelectrodes with two different heights for both neural stimulation and recording

Microelectrode array (MEA) is one of the representative platforms for recording and stimulating neurons to explore their populational electrophysiological activities. Due to the layered structures of the nervous system, various three-dimensional (3D) MEAs have been used to reach specific target depths of neuronal tissues. But it is still difficult to monolithically fabricate various heights of microelectrodes, particularly with optical transparency. Here, we present a 3D MEA with different heights fabricated by multi-step silicon deep reactive ion etching. The microelectrodes were embedded in reflowed glass for high transparency which enables both neural tissue observation and optical stimulation. The heights of microelectrodes were well controlled by our fabrication processes and electrical connections were made with through glass vias (TGVs) to the bottom side for individual addressing. To confine electric stimulation in a small area, we used microelectrodes neighboring a stimulation electrode as local returns. The feasibility of the fabricated MEA was demonstrated by stimulating the mouse retina and successfully recording spikes of ganglion cells located near the stimulation site. Consistent with our COMSOL simulation, the ex vivo experimental results showed the stimulation current confinement was effective. Our study provides a new tool for 3D nervous systems and its fabrication method.

Closed-loop parameter optimization for patient-specific phrenic nerve stimulation.

Ventilator-induced diaphragm dysfunction occurs rapidly following the onset of mechanical ventilation and has significant clinical consequences. Phrenic nerve stimulation has shown promise in maintaining diaphragm function by inducing diaphragm contractions. Non-invasive stimulation is an attractive option as it minimizes the procedural risks associated with invasive approaches. However, this method is limited by sensitivity to electrode position and inter-individual variability in stimulation thresholds. This makes clinical application challenging due to potentially time-consuming calibration processes to achieve reliable stimulation. We applied non-invasive electrical stimulation to the phrenic nerve in the neck in healthy volunteers. A closed-loop system recorded the respiratory flow produced by stimulation and automatically adjusted the electrode position and stimulation amplitude based on the respiratory response. By iterating over electrodes, the optimal electrode was selected. A binary search method over stimulation amplitudes was then employed to determine an individualized stimulation threshold. Pulse trains above this threshold were delivered to produce diaphragm contraction. Nine healthy volunteers were recruited. Mean threshold stimulation amplitude was 36.17 ± 14.34 mA (range 19.38-59.06 mA). The threshold amplitude for reliable nerve capture was moderately correlated with BMI (Pearson's r = 0.66, p = 0.049). Repeating threshold measurements within subjects demonstrated low intra-subject variability of 2.15 ± 1.61 mA between maximum and minimum thresholds on repeated trials. Bilateral stimulation with individually optimized parameters generated reliable diaphragm contraction, resulting in significant inhaled volumes following stimulation. We demonstrate the feasibility of a system for automatic optimization of electrode position and stimulation parameters using a closed-loop system. This opens the possibility of easily deployable individualized stimulation in the intensive care setting to reduce ventilator-induced diaphragm dysfunction.

A Wireless, Mechanically Flexible, 25μm-Thick, 65,536-Channel Subdural Surface Recording and Stimulating Microelectrode Array with Integrated Antennas.

This paper presents a fully wireless microelectrode array (MEA) system-on-chip (SoC) with 65,536 electrodes for non-penetrative cortical recording and stimulation, featuring a total sensing area of 6.8mm×7.4mm with a 26.5μm×29μm electrode pitch. Sensing, data telemetry, and powering are monolithically integrated on a single chip, which is made mechanically flexible to conform to the surface of the brain by substrate removal to a total thickness of 25μm allowing it to be contained entirely in the subdural space under the skull.

Placement of Extracranial Stimulating Electrodes and Measurement of Cerebral Blood Flow and Intracranial Electrical Fields in Anesthetized Mice.

The detection of cerebral blood flow (CBF) responses to various forms of neuronal activation is critical for understanding dynamic brain function and variations in the substrate supply to the brain. This paper describes a protocol for measuring CBF responses to transcranial alternating current stimulation (tACS). Dose-response curves are estimated both from the CBF change occurring with tACS (mA) and from the intracranial electric field (mV/mm). We estimate the intracranial electrical field based on the different amplitudes measured by glass microelectrodes within each side of the brain. In this paper, we describe the experimental setup, which involves using either bilateral laser Doppler (LD) probes or laser speckle imaging (LSI) to measure the CBF; as a result, this setup requires anesthesia for the electrode placement and stability. We present a correlation between the CBF response and the current as a function of age, showing a significantly larger response at higher currents (1.5 mA and 2.0 mA) in young control animals (12-14 weeks) compared to older animals (28-32 weeks) (p < 0.005 difference). We also demonstrate a significant CBF response at electrical field strengths <5 mV/mm, which is an important consideration for eventual human studies. These CBF responses are also strongly influenced by the use of anesthesia compared to awake animals, the respiration control (i.e., intubated vs. spontaneous breathing), systemic factors (i.e., CO2), and local conduction within the blood vessels, which is mediated by pericytes and endothelial cells. Likewise, more detailed imaging/recording techniques may limit the field size from the entire brain to only a small region. We describe the use of extracranial electrodes for applying tACS stimulation, including both homemade and commercial electrode designs for rodents, the concurrent measurement of the CBF and intracranial electrical field using bilateral glass DC recording electrodes, and the imaging approaches. We are currently applying these techniques to implement a closed-loop format for augmenting the CBF in animal models of Alzheimer's disease and stroke.