Infrared Neural Stimulation Research Articles

Infrared neuroglial modulation of spinal locomotor networks

Infrared neural stimulation (INS) emerges as a promising tool for stimulating the nervous system by its high spatial precision and absence of the use of exogenous agents into the tissue, which led to the first successful proof of concept in human brain. While neural networks have been the focal point of INS research, this technique is also non cell type specific as it triggers activity in non electrically excitable cells. Despite increasing interest, there remains to demonstrate well defined simultaneous astrocytic and neuronal signals in response to INS. Using calcium imaging, we show that INS has the capacity to initiate calcium signaling in both astrocytes and neurons simultaneously from the rostral lumbar spinal cord, each exhibiting distinct temporal and amplitude characteristics. Importantly, the mechanism underlying infrared-induced neuronal and astrocytic calcium signaling differ, with neuronal activity relying on sodium channels, whereas induced astrocytic signaling is predominantly influenced by extracellular calcium and TRPV4 channels. Furthermore, our findings demonstrate the frequency shift of neuronal calcium oscillations through infrared stimulation. By deepening our understanding in INS fundamentals, this technique holds great promise for advancing neuroscience, deepening our understanding of pathologies, and potentially paving the way for future clinical applications.

Brainwide mesoscale functional networks revealed by focal infrared neural stimulation of the amygdala.

The primate amygdala serves to evaluate emotional content of sensory inputs and modulate emotional and social behaviors; it modulates cognitive, multisensory and autonomic circuits predominantly via the basal (BA), lateral (LA), and central (CeA) nuclei, respectively. Based on recent electrophysiological evidence suggesting mesoscale (millimeters-scale) nature of intra-amygdala functional organization, we have investigated the connectivity of these nuclei using Infrared Neural Stimulation of single mesoscale sites coupled with mapping in ultrahigh field 7T functional Magnetic Resonance Imaging (INS-fMRI). Stimulation of multiple sites within amygdala of single individuals evoked 'mesoscale functional connectivity maps', allowing comparison of BA, LA and CeA connected brainwide networks. This revealed a mesoscale nature of connected sites, complementary spatial patterns of functional connectivity, and topographic relationships of nucleus-specific connections. Our data reveal a functional architecture of systematically organized brainwide networks mediating sensory, cognitive, and autonomic influences from the amygdala.

Quantifying tissue temperature changes induced by infrared neural stimulation: numerical simulation and MR thermometry.

Infrared neural stimulation (INS) delivered via short pulse trains is an innovative tool that has potential for us use for studying brain function and circuitry, brain machine interface, and clinical use. The prevailing mechanism for INS involves the conversion of light energy into thermal transients, leading to neuronal membrane depolarization. Due to the potential risks of thermal damage, it is crucial to ensure that the resulting local temperature increases are within non-damaging limits for brain tissues. Previous studies have estimated damage thresholds using histological methods and have modeled thermal effects based on peripheral nerves. However, additional quantitative measurements and modeling studies are needed for the central nervous system. Here, we performed 7 T MRI thermometry on ex vivo rat brains following the delivery of infrared pulse trains at five different intensities from 0.1-1.0 J/cm2 (each pulse train 1,875 nm, 25 us/pulse, 200 Hz, 0.5 s duration, delivered through 200 µm fiber). Additionally, we utilized the General BioHeat Transfer Model (GBHTM) to simulate local temperature changes in perfused brain tissues while delivering these laser energies to tissue (with optical parameters of human skin) via three different sizes of optical fibers at five energy intensities. The simulation results clearly demonstrate that a 0.5 second INS pulse train induces an increase followed by an immediate drop in temperature at stimulation offset. The delivery of multiple pulse trains with 2.5 s interstimulus interval (ISI) leads to rising temperatures that plateau. Both thermometry and modeling results show that, using parameters that are commonly used in biological applications (200 µm diameter fiber, 0.1-1.0 J/cm2), the final temperature increase at the end of the 60 sec stimuli duration does not exceed 1°C with stimulation values of 0.1-0.5 J/cm2 and does not exceed 2°C with stimulation values of up to 1.0 J/cm2. Thus, the maximum temperature rise is consistent with the thermal damage threshold reported in previous studies. This study provides a quantitative evaluation of the temperature changes induced by INS, suggesting that existing practices pose minimal major safety concerns for biological tissues.

Modulatory Effects on Laminar Neural Activity Induced by Near-Infrared Light Stimulation with a Continuous Waveform to the Mouse Inferior Colliculus In Vivo.

Infrared neural stimulation (INS) is a promising area of interest for the clinical application of a neuromodulation method. This is in part because of its low invasiveness, whereby INS modulates the activity of the neural tissue mainly through temperature changes. Additionally, INS may provide localized brain stimulation with less tissue damage. The inferior colliculus (IC) is a crucial auditory relay nucleus and a potential target for clinical application of INS to treat auditory diseases and develop artificial hearing devices. Here, using continuous INS with low to high-power density, we demonstrate the laminar modulation of neural activity in the mouse IC in the presence and absence of sound. We investigated stimulation parameters of INS to effectively modulate the neural activity in a facilitatory or inhibitory manner. A mathematical model of INS-driven brain tissue was first simulated, temperature distributions were numerically estimated, and stimulus parameters were selected from the simulation results. Subsequently, INS was administered to the IC of anesthetized mice, and the modulation effect on the neural activity was measured using an electrophysiological approach. We found that the modulatory effect of INS on the spontaneous neural activity was bidirectional between facilitatory and inhibitory effects. The modulatory effect on sound-evoked responses produced only an inhibitory effect to all examined stimulus intensities. Thus, this study provides important physiological evidence on the response properties of IC neurons to INS. Overall, INS can be used for the development of new therapies for neurological disorders and functional support devices for auditory central processing.

Characterization and closed-loop control of infrared thalamocortical stimulation produces spatially constrained single-unit responses.

Deep brain stimulation (DBS) is a powerful tool for the treatment of circuitopathy-related neurological and psychiatric diseases and disorders such as Parkinson's disease and obsessive-compulsive disorder, as well as a critical research tool for perturbing neural circuits and exploring neuroprostheses. Electrically mediated DBS, however, is limited by the spread of stimulus currents into tissue unrelated to disease course and treatment, potentially causing undesirable patient side effects. In this work, we utilize infrared neural stimulation (INS), an optical neuromodulation technique that uses near to midinfrared light to drive graded excitatory and inhibitory responses in nerves and neurons, to facilitate an optical and spatially constrained DBS paradigm. INS has been shown to provide spatially constrained responses in cortical neurons and, unlike other optical techniques, does not require genetic modification of the neural target. We show that INS produces graded, biophysically relevant single-unit responses with robust information transfer in rat thalamocortical circuits. Importantly, we show that cortical spread of activation from thalamic INS produces more spatially constrained response profiles than conventional electrical stimulation. Owing to observed spatial precision of INS, we used deep reinforcement learning (RL) for closed-loop control of thalamocortical circuits, creating real-time representations of stimulus-response dynamics while driving cortical neurons to precise firing patterns. Our data suggest that INS can serve as a targeted and dynamic stimulation paradigm for both open and closed-loop DBS.

Heat accumulation during infrared stimulation impacts the response of ex vivo rat sciatic nerve.

Infrared neural stimulation (INS) is a neuromodulation technique that involves short optical pulses delivered to the neural tissue, resulting in the initiation of action potentials. In this work, we studied the compound neural action potentials (CNAP) generated by INS in five ex vivo sciatic nerves. A 1470 nm laser emitting a sequence of 0.4 ms light pulses with a peak power of 10 W was used. A single 4 mJ stimulus is not capable of eliciting a nerve response. However, repetition of the optical stimuli resulted in the induction of CNAPs. Heat accumulation induced by repetition rates as high as 10 Hz may be involved in the increase in CNAP amplitude. This sensitization effect may help to reduce the pulse energy required to evoke CNAP. In addition, these results highlight the importance of investigating the role of the slow nerve temperature dynamics in INS.

Design and application of a multimodality-compatible 1Tx/6Rx RF coil for monkey brain MRI at 7T

ObjectiveBlood-oxygen-level-dependent functional MRI allows to investigte neural activities and connectivity. While the non-human primate plays an essential role in neuroscience research, multimodal methods combining functional MRI with other neuroimaging and neuromodulation enable us to understand the brain network at multiple scales. ApproachIn this study, a tight-fitting helmet-shape receive array with a single transmit loop for anesthetized macaque brain MRI at 7T was fabricated with four openings constructed in the coil housing to accommodate multimodal devices, and the coil performance was quantitatively evaluated and compared to a commercial knee coil. In addition, experiments over three macaques with infrared neural stimulation (INS), focused ultrasound stimulation (FUS), and transcranial direct current stimulation (tDCS) were conducted. Main resultsThe RF coil showed higher transmit efficiency, comparable homogeneity, improved SNR and enlarged signal coverage over the macaque brain. Infrared neural stimulation was applied to the amygdala in deep brain region, and activations in stimulation sites and connected sites were detected, with the connectivity consistent with anatomical information. Focused ultrasound stimulation was applied to the left visual cortex, and activations were acquired along the ultrasound traveling path, with all time course curves consistent with pre-designed paradigms. The existence of transcranial direct current stimulation electrodes brought no interference to the RF system, as evidenced through high-resolution MPRAGE structure images. SignificanceThis pilot study reveals the feasibility for brain investigation at multiple spatiotemporal scales, which may advance our understanding in dynamic brain networks.

Infrared neural stimulation markedly enhances nerve functionality assessment during nerve monitoring

In surgical procedures where the risk of accidental nerve damage is prevalent, surgeons commonly use electrical stimulation (ES) during intraoperative nerve monitoring (IONM) to assess a nerve’s functional integrity. ES, however, is subject to off-target stimulation and stimulation artifacts disguising the true functionality of the specific target and complicating interpretation. Lacking a stimulation artifact and having a higher degree of spatial specificity, infrared neural stimulation (INS) has the potential to improve upon clinical ES for IONM. Here, we present a direct comparison between clinical ES and INS for IONM performance in an in vivo rat model. The sensitivity of INS surpasses that of ES in detecting partial forms of damage while maintaining a comparable specificity and sensitivity to more complete forms. Without loss in performance, INS is readily compatible with existing clinical nerve monitoring systems. These findings underscore the clinical potential of INS to improve IONM and surgical outcomes.

Infrared neural stimulation in human cerebral cortex

BackgroundModulation of brain circuits by electrical stimulation has led to exciting and powerful therapies for diseases such as Parkinson's. Because human brain organization is based in mesoscale (millimeter-scale) functional nodes, having a method that can selectively target such nodes could enable more precise, functionally specific stimulation therapies. Infrared Neural Stimulation (INS) is an emerging stimulation technology that stimulates neural tissue via delivery of tiny heat pulses. In nonhuman primates, this optical method provides focal intensity-dependent stimulation of the brain without tissue damage. However, whether INS application to the human central nervous system (CNS) is similarly effective is unknown. ObjectiveTo examine the effectiveness of INS on human cerebral cortex in intraoperative setting and to evaluate INS damage threshholds. MethodsFive epileptic subjects undergoing standard lobectomy for epilepsy consented to this study. Cortical response to INS was assessed by intrinsic signal optical imaging (OI, a method that detects changes in tissue reflectance due to neuronal activity). A custom integrated INS and OI system was developed specifically for short-duration INS and OI acquisition during surgical procedures. Single pulse trains of INS with intensities from 0.2 to 0.8 J/cm2 were delivered to the somatosensory cortex and responses were recorded via optical imaging. Following tissue resection, histological analysis was conducted to evaluate damage threshholds. ResultsAs assessed by OI, and similar to results in monkeys, INS induced responses in human cortex were highly focal (millimeter sized) and led to relative suppression of nearby cortical sites. Intensity dependence was observed at both stimulated and functionally connected sites. Histological analysis of INS-stimulated human cortical tissue provided damage threshold estimates. ConclusionThis is the first study demonstrating application of INS to human CNS and shows feasibility for stimulating single cortical nodes and associated sites and provided INS damage threshold estimates for cortical tissue. Our results suggest that INS is a promising tool for stimulation of functionally selective mesoscale circuits in the human brain, and may lead to advances in the future of precision medicine.

Response of a neuronal network computational model to infrared neural stimulation.

Infrared neural stimulation (INS), as a novel form of neuromodulation, allows modulating the activity of nerve cells through thermally induced capacitive currents and thermal sensitivity ion channels. However, fundamental questions remain about the exact mechanism of INS and how the photothermal effect influences the neural response. Computational neural modeling can provide a powerful methodology for understanding the law of action of INS. We developed a temperature-dependent model of ion channels and membrane capacitance based on the photothermal effect to quantify the effect of INS on the direct response of individual neurons and neuronal networks. The neurons were connected through excitatory and inhibitory synapses and constituted a complex neuronal network model. Our results showed that a slight increase in temperature promoted the neuronal spikes and enhanced network activity, whereas the ultra-temperature inhibited neuronal activity. This biophysically based simulation illustrated the optical dose-dependent biphasic cell response with capacitive current as the core change condition. The computational model provided a new sight to elucidate mechanisms and inform parameter selection of INS.

Visualizing the lipid dynamics role in infrared neural stimulation using stimulated Raman scattering

Infrared neural stimulation (INS) uses pulsed infrared light to yield label-free neural stimulation with broad experimental and translational utility. Despite its robust demonstration, INS’s mechanistic and biophysical underpinnings have been the subject of debate for more than a decade. The role of lipid membrane thermodynamics appears to play an important role in how fast IR-mediated heating nonspecifically drives action potential generation. Direct observation of lipid membrane dynamics during INS remains to be shown in a live neural model system. We used hyperspectral stimulated Raman scattering microscopy to study biochemical signatures of high-speed vibrational dynamics underlying INS in a live neural cell culture model. The findings suggest that lipid bilayer structural changes occur during INS in vitro in NG108-15 neuroglioma cells. Lipid-specific signatures of cell stimulated Raman scattering spectra varied with stimulation energy and radiation exposure. The spectroscopic observations agree with high-speed ratiometric fluorescence imaging of a conventional lipophilic membrane structure reporter, 4-(2-(6-(dibutylamino)-2-naphthalenyl)ethenyl)-1-(3-sulfopropyl)pyridinium hydroxide. The findings support the hypothesis that INS causes changes in the lipid membrane of neural cells by changing the lipid membrane packing order. This work highlights the potential of hyperspectral stimulated Raman scattering as a method to safely study biophysical and biochemical dynamics in live cells.

Fiberoptic array for multiple channel infrared neural stimulation of the brain.

.Significance: We present a new optical method for modulating cortical activity in multiple locations and across multiple time points with high spatial and temporal precision. Our method uses infrared light and does not require dyes or transgenic modifications. It is compatible with a number of other stimulation and recording techniques.Aim: Infrared neural stimulation (INS) has been largely confined to single point stimuli. In this study, we expand upon this approach and develop a rapidly switched fiber array capable of generation of stimulus patterns. Our prototype is capable of stimulating at nine separate locations but is easily scalable.Approach: Our device is made of commercially available components: a solid-state infrared laser, a piezoelectric fiber coupled optical switch, and diameter optical fibers. We validate it using intrinsic optical signal imaging of INS responses in macaque and squirrel monkey sensory cortical areas.Results: We demonstrate that our switched array can consistently generate responses in primate cortex, consistent with earlier single channel INS investigations.Conclusions: Our device can successfully target the cortical surface, either at one specific region or multiple points spread out across different areas. It is compatible with a host of other imaging and stimulation modalities.

Identifying optimal parameters for infrared neural stimulation in the peripheral nervous system.

.Significance: Infrared neural stimulation (INS) utilizes pulsed infrared light to selectively elicit neural activity without exogenous compounds. Despite its versatility in a broad range of biomedical applications, no comprehensive comparison of factors pertaining to the efficacy and safety of INS such as wavelength, radiant exposure, and optical spot size exists in the literature.Aim: Here, we evaluate these parameters using three of the wavelengths commonly used for INS, 1450 nm, 1875 nm, and 2120 nm.Approach: In an in vivo rat sciatic nerve preparation, the stimulation threshold and transition rate to 100% activation probability were used to compare the effects of each parameter.Results: The pulsed diode lasers at 1450 nm and 1875 nm had a consistently higher () stimulation threshold than that of the Ho:YAG laser at 2120 nm (). In addition, the Ho:YAG produced a faster transition rate to 100% activation probability compared to the diode lasers. Our data suggest that the superior performance of the Ho:YAG is a result of the high-intensity microsecond spike at the onset of the pulse. Acute histological evaluation of diode irradiated nerves revealed a safe range of radiant exposures for stimulation.Conclusion: Together, our results identify measures to improve the safety, efficacy, and accessibility of INS technology for research and clinical applications.

Short-wave Infrared Neural Stimulation Drives Graded Sciatic Nerve Activation Across A Continuum of Wavelengths.

Infrared neural stimulation (INS) is an optical stimulation technique which uses coherent light to stimulate nerves and neurons and which shows increased spatial selectivity compared to electrical stimulation. This could improve deep brain, high channel count, or vagus nerve stimulation. In this study, we seek to understand the wavelength dependence of INS in the near-infrared optical window. Rat sciatic nerves were excised ex vivo and stimulated with wavelengths between 700 and 900 nm. Recorded compound nerve action potentials (CNAPs) showed that stimulation was maximized in the 700 nm window despite comparable laser power levels across wavelengths. Computational models demonstrated that wavelength-based activation dependencies were not a result of passive optical properties. This data demonstrates that INS is both wavelength and power level dependent, which inform stimulation systems to actively target neural microcircuits in humans.

Thermal damage threshold of neurons during infrared stimulation.

In infrared neural stimulation (INS), laser-evoked thermal transients are used to generate small depolarising currents in neurons. The laser exposure poses a moderate risk of thermal damage to the target neuron. Indeed, exogenous methods of neural stimulation often place the target neurons under stressful non-physiological conditions, which can hinder ordinary neuronal function and hasten cell death. Therefore, quantifying the exposure-dependent probability of neuronal damage is essential for identifying safe operating limits of INS and other interventions for therapeutic and prosthetic use. Using patch-clamp recordings in isolated spiral ganglion neurons, we describe a method for determining the dose-dependent damage probabilities of individual neurons in response to both acute and cumulative infrared exposure parameters based on changes in injection current. The results identify a local thermal damage threshold at approximately 60 °C, which is in keeping with previous literature and supports the claim that damage during INS is a purely thermal phenomenon. In principle this method can be applied to any potentially injurious stimuli, allowing for the calculation of a wide range of dose-dependent neural damage probabilities. Unlike histological analyses, the technique is well-suited to quantifying gradual neuronal damage, and critical threshold behaviour is not required.

Blind Localization of Heating in Neural Tissues Induced by a Train of the Infrared Pulse Laser.

Introduction: Recently, infrared lasers (wavelengths larger than 1100 nm) have been applied to stimulate neural tissues. Infrared neural stimulation (INS) has some advantages over conventional electric stimulation, including contact-free delivery, spatial precision, and lack of stimulation artifacts. In this study and based on a photothermal mechanism, we applied the heat diffusion equation to study temperature variation of a biological phantom during INS. In addition, the impact of laser parameters on spatially localized heating induced by 2 different infrared wavelengths were studied. Methods: We studied the localization of INS inside a phantom similar to cortical neural tissue. First, we analytically solved the heat diffusion equation to study the distribution of temperature inside this phantom. Then, the accuracy of analytical results was verified by heating the phantom using amplitude-modulated infrared lasers (lambda= 1450 and 1500 nm, the energy between 2 and 5 mJ and pulse duration up to 20 ms). The laser light was directed to sample by a multimode optical fiber (NA=0.22, core size= 200 microns). Finally, the impacts of laser properties on the spatial resolution of infrared heating were discerned. Results: In order to verify analytical results, we measured the maximum temperatures of the phantom during illumination of lasers and compared them with analytical results. The analytical results were in agreement with the experimental results. The effects of laser beam properties such as pulse duration, energy and repetition rate frequency on the spatial resolution were investigated. The results indicated that the spatial resolution of INS can be smaller than one millimeter. Conclusion: Here, the influences of laser properties on the localization of INS inside a biological phantom were studied. These results can be applied to improve the spatial selectivity of the peripheral nerve interface.

Gold nanorod-assisted near-infrared stimulation of bullfrog sciatic nerve.

Infrared neural stimulation (INS) is a new and developing approach for neural repair, with the advantages of being non-contact, spatially precise, and artifact-free. However, the disadvantage of infrared light is that it is difficult to stimulate deep tissue because of its weak penetrating power. Therefore, this paper introduces an improved method using near-infrared laser to stimulate bullfrog sciatic nerves because of its strong penetrating power. Meanwhile, gold nanorods (Au NRs) are injected into the nerve to increase the absorption of light. The mechanism is the instantaneous temperature rise caused by the absorption of infrared light by Au NRs. The compound muscle action potential (CMAP) associated with the irradiated sciatic nerve is recorded by a multi-channel physiological signal instrument. The peak to peak amplitude (Vpp) of CMAP for sciatic nerves injected with Au NRs increases significantly compared to the CMAP for control nerves without Au NRs. These results demonstrate INS by labeling nerves with nanoparticle exhibiting latent capacity to increase the efficiency, spatial resolution, and the neural responsivity, and especially, can increase the penetration depth and reduce the requisite radiant exposure level.

A multimodal microtool for spatially controlled infrared neural stimulation in the deep brain tissue

Infrared neural stimulation (INS) uses pulsed mid-infrared light to generate highly controlled temperature transients in neurons, leading them to fire action potentials. Stimulation of the superficial layer of the intact brain has been demonstrated, however, intervention in the deep neural tissue has larger potential in view of therapeutic use. To reveal the underlying mechanism of deep tissue stimulation properly, we present the design, the fabrication scheme and functional testing of a novel, multimodal optrode for future INS experiments in vivo. Three modalities – electrophysiological recording, thermal measurement and delivery of infrared light – were integrated using silicon MEMS technology. The average overall efficiency of the microoptical system delivering the infrared light at chip-scale is measured as 32.04 ± 4.10%, while the max. efficiency in packaged form is 41.5 ± 3.29%. The average beam spot size at the probe tip is 0.024 ± 0.006 mm2. The temperature coefficient of resistance of the integrated thermal sensor monitoring the change in background temperature is 2636 ± 75 ppm/°C. The average impedance of the electrophysiological recording sites is 1031 ± 95 kΩ. Based on beam profile measurements and multiphysical modeling, the spatial extent of the thermally excited region and its temporal dynamics are estimated in case of both low (30 mW) and high (3 W) power excitation. Due to the monolithically integrated functionalities, a single probe is sufficient to determine safe stimulation parameters in vivo. As far as we know, this is the first planar, multimodal optrode designed for INS studies in the deep tissue.

Auditory Neural Activity in Congenitally Deaf Mice Induced by Infrared Neural Stimulation

To determine whether responses during infrared neural stimulation (INS) result from the direct interaction with spiral ganglion neurons (SGNs), we tested three genetically modified deaf mouse models: Atoh1-cre; Atoh1f/f (Atoh1 conditional knockout, CKO), Atoh1-cre; Atoh1f/kiNeurog1 (Neurog1 knockin, KI), and the Vglut3 knockout (Vglut3−/−) mice. All animals were exposed to tone bursts and clicks up to 107 dB (re 20 µPa) and to INS, delivered with a 200 µm optical fiber. The wavelength (λ) was 1860 nm, the radiant energy (Q) 0-800 µJ/pulse, and the pulse width (PW) 100–500 µs. No auditory responses to acoustic stimuli could be evoked in any of these animals. INS could not evoke auditory brainstem responses in Atoh1 CKO mice but could in Neurog1 KI and Vglut3−/− mice. X-ray micro-computed tomography of the cochleae showed that responses correlated with the presence of SGNs and hair cells. Results in Neurog1 KI mice do not support a mechanical stimulation through the vibration of the basilar membrane, but cannot rule out the direct activation of the inner hair cells. Results in Vglut3−/− mice, which have no synaptic transmission between inner hair cells and SGNs, suggested that hair cells are not required.

Model study of combined electrical and near-infrared neural stimulation on the bullfrog sciatic nerve.

This paper implemented a model study of combined electrical and near-infrared (808nm) neural stimulation (NINS) on the bullfrog sciatic nerve. The model includes a COMSOL model to calculate the electric-field distribution of the surrounding area of the nerve, a Monte Carlo model to simulate light transport and absorption in the bullfrog sciatic nerve during NINS, and a NEURON model to simulate the neural electrophysiology changes under electrical stimulus and laser irradiation. The optical thermal effect is considered the main mechanism during NINS. Therefore, thermal change during laser irradiation was calculated by the Monte Carlo method, and the temperature distribution was then transferred to the NEURON model to stimulate the sciatic nerve. The effects on thermal response by adjusting the laser spot size, energy of the beam, and the absorption coefficient of the nerve are analyzed. The effect of the ambient temperature on the electrical stimulation or laser stimulation and the interaction between laser irradiation and electrical stimulation are also studied. The results indicate that the needed stimulus threshold for neural activation or inhibition is reduced by laser irradiation. Additionally, the needed laser energy for blocking the action potential is reduced by electrical stimulus. Both electrical and laser stimulation are affected by the ambient temperature. These results provide references for subsequent animal experiments and could be of great help to future basic and applied studies of infrared neural stimulation (INS).