Long-term intravital subcellular imaging with confocal scanning light-field microscopy.

Long-term observation of subcellular dynamics in living organisms is limited by background fluorescence originating from tissue scattering or dense labeling. Existing confocal approaches face an inevitable tradeoff among parallelization, resolution and phototoxicity. Here we present confocal scanning light-field microscopy (csLFM), which integrates axially elongated line-confocal illumination with the rolling shutter in scanning light-field microscopy (sLFM). csLFM enables high-fidelity, high-speed, three-dimensional (3D) imaging at near-diffraction-limit resolution with both optical sectioning and low phototoxicity. By simultaneous 3D excitation and detection, the excitation intensity can be reduced below 1 mW mm-2, with 15-fold higher signal-to-background ratio over sLFM. We imaged subcellular dynamics over 25,000 timeframes in optically challenging environments in different species, such as migrasome delivery in mouse spleen, retractosome generation in mouse liver and 3D voltage imaging in Drosophila. Moreover, csLFM facilitates high-fidelity, large-scale neural recording with reduced crosstalk, leading to high orientation selectivity to visual stimuli, similar to two-photon microscopy, which aids understanding of neural coding mechanisms.

Distinct neural activities of the cortical layer 2/3 across isoflurane anesthesia: A large-scale simultaneous observation of neurons.

Anesthesia inhibits neural activity in the brain, causing patients to lose consciousness and sensation during the surgery. Layers 2/3 of the cortex are important structures for the integration of information and consciousness, which are closely related to normal cognitive function. However, the dynamics of the large-scale population of neurons across multiple regions in layer 2/3 during anesthesia and recovery processes remains unclear. We conducted simultaneous observations and analysis of large-scale calcium signaling dynamics across multiple cortical regions within cortical layer 2/3 during isoflurane anesthesia and recovery in vivo by high-resolution wide-field microscopy. Under isoflurane-induced anesthesia, there is an overall decrease in neuronal activity across multiple regions in the cortical layer 2/3. Notably, some neurons display a paradoxical increase in activity during anesthesia. Additionally, the activity among multiple cortical regions under anesthesia was homogeneous. It is only during the recovery phase that variability emerges in the extent of increased neural activity across different cortical regions. Within the same duration of anesthesia, neural activity did not return to preanesthetic levels. To sum up, anesthesia as a dynamic alteration of brain functional networks, encompassing shifts in patterns of neural activity, homogeneousness among cortical neurons and regions, and changes in functional connectivity. Recovery from anesthesia does not entail a reversal of these effects within the same timeframe.

Multifocal fluorescence video-rate imaging of centimetre-wide arbitrarily shaped brain surfaces at micrometric resolution.

Fluorescence microscopy allows for the high-throughput imaging of cellular activity across brain areas in mammals. However, capturing rapid cellular dynamics across the curved cortical surface is challenging, owing to trade-offs in image resolution, speed, field of view and depth of field. Here we report a technique for wide-field fluorescence imaging that leverages selective illumination and the integration of focal areas at different depths via a spinning disc with varying thickness to enable video-rate imaging of previously reconstructed centimetre-scale arbitrarily shaped surfaces at micrometre-scale resolution and at a depth of field of millimetres. By implementing the technique in a microscope capable of acquiring images at 1.68 billion pixels per second and resolving 16.8 billion voxels per second, we recorded neural activities and the trajectories of neutrophils in real time on curved cortical surfaces in live mice. The technique can be integrated into many microscopes and macroscopes, in both reflective and fluorescence modes, for the study of multiscale cellular interactions on arbitrarily shaped surfaces.

Neuron populations across layer 2-6 in the mouse visual cortex exhibit different coding abilities in the awake mice.

Introduction: The visual cortex is a key region in the mouse brain, responsible for processing visual information. Comprised of six distinct layers, each with unique neuronal types and connections, the visual cortex exhibits diverse decoding properties across its layers. This study aimed to investigate the relationship between visual stimulus decoding properties and the cortical layers of the visual cortex while considering how this relationship varies across different decoders and brain regions. Methods: This study reached the above conclusions by analyzing two publicly available datasets obtained through two-photon microscopy of visual cortex neuronal responses. Various types of decoders were tested for visual cortex decoding. Results: Our findings indicate that the decoding accuracy of neuronal populations with consistent sizes varies among visual cortical layers for visual stimuli such as drift gratings and natural images. In particular, layer 4 neurons in VISp exhibited significantly higher decoding accuracy for visual stimulus identity compared to other layers. However, in VISm, the decoding accuracy of neuronal populations with the same size in layer 2/3 was higher than that in layer 4, despite the overall accuracy being lower than that in VISp and VISl. Furthermore, SVM surpassed other decoders in terms of accuracy, with the variation in decoding performance across layers being consistent among decoders. Additionally, we found that the difference in decoding accuracy across different imaging depths was not associated with the mean orientation selectivity index (OSI) and the mean direction selectivity index (DSI) neurons, but showed a significant positive correlation with the mean reliability and mean signal-to-noise ratio (SNR) of each layer's neuron population. Discussion: These findings lend new insights into the decoding properties of the visual cortex, highlighting the role of different cortical layers and decoders in determining decoding accuracy. The correlations identified between decoding accuracy and factors such as reliability and SNR pave the way for more nuanced understandings of visual cortex functioning.

Cerebral cortex and hippocampus neural interaction during vagus nerve stimulation under in vivo large-scale imaging.

Objective: The purpose of this study was to study mechanisms of VNS modulation from a single neuron perspective utilizing a practical observation platform with single neuron resolution and widefield, real-time imaging coupled with an animal model simultaneously exposing the cerebral cortex and the hippocampus. Methods: We utilized the observation platform characterized of widefield of view, real-time imaging, and high spatiotemporal resolution to obtain the neuronal activities in the cerebral cortex and the hippocampus during VNS in awake states and under anesthesia. Results: Some neurons in the hippocampus were tightly related to VNS modulation, and varied types of neurons showed distinct responses to VNS modulation. Conclusion: We utilized such an observation platform coupled with a novel animal model to obtain more information on neuron activities in the cerebral cortex and the hippocampus, providing an effective method to further study the mechanisms of therapeutic effects modulated by VNS.

Rapid detection of neurons in widefield calcium imaging datasets after training with synthetic data.

Widefield microscopy can provide optical access to multi-millimeter fields of view and thousands of neurons in mammalian brains at video rate. However, tissue scattering and background contamination results in signal deterioration, making the extraction of neuronal activity challenging, laborious and time consuming. Here we present our deep-learning-based widefield neuron finder (DeepWonder), which is trained by simulated functional recordings and effectively works on experimental data to achieve high-fidelity neuronal extraction. Equipped with systematic background contribution priors, DeepWonder conducts neuronal inference with an order-of-magnitude-faster speed and improved accuracy compared with alternative approaches. DeepWonder removes background contaminations and is computationally efficient. Specifically, DeepWonder accomplishes 50-fold signal-to-background ratio enhancement when processing terabytes-scale cortex-wide functional recordings, with over 14,000 neurons extracted in 17 h.

Real-time denoising enables high-sensitivity fluorescence time-lapse imaging beyond the shot-noise limit.

A fundamental challenge in fluorescence microscopy is the photon shot noise arising from the inevitable stochasticity of photon detection. Noise increases measurement uncertainty and limits imaging resolution, speed and sensitivity. To achieve high-sensitivity fluorescence imaging beyond the shot-noise limit, we present DeepCAD-RT, a self-supervised deep learning method for real-time noise suppression. Based on our previous framework DeepCAD, we reduced the number of network parameters by 94%, memory consumption by 27-fold and processing time by a factor of 20, allowing real-time processing on a two-photon microscope. A high imaging signal-to-noise ratio can be acquired with tenfold fewer photons than in standard imaging approaches. We demonstrate the utility of DeepCAD-RT in a series of photon-limited experiments, including in vivo calcium imaging of mice, zebrafish larva and fruit flies, recording of three-dimensional (3D) migration of neutrophils after acute brain injury and imaging of 3D dynamics of cortical ATP release. DeepCAD-RT will facilitate the morphological and functional interrogation of biological dynamics with a minimal photon budget.

The burst of electrophysiological signals in the suprachiasmatic nucleus of mouse during the arousal detected by microelectrode arrays.

The neural mechanisms of torpor have essential reference significance for medical methods and long-term manned space. Changes in electrophysiology of suprachiasmatic nucleus (SCN) conduce to revealing the neural mechanisms from the torpor to arousal. Due to the lower physiology state during the torpor, it is a challenge to detect neural activities in vivo on freely behaving mice. Here, we introduced a multichannel microelectrode array (MEA) for real-time detection of local field potential (LFP) and action potential (spike) in the SCN in induced torpor mice. Meanwhile, core body temperature and behaviors of mice were recorded for further analysis. Platinum nanoparticles (PtNPs) and Nafion membrane modified MEA has a lower impedance (16.58 ± 3.93 kΩ) and higher signal-to-noise ratio (S/N = 6.1). We found that from torpor to arousal, the proportion of theta frequency bands of LFPs increased, spike firing rates rapidly increased. These results could all be characteristic information of arousal, supported by the microscopic neural activity promoting arousal in mice. MEA displayed real-time dynamic changes of neuronal activities in the SCN, which was more helpful to analyze and understand neural mechanisms of torpor and arousal. Our study provided a factual basis for the neural state in SCN of induced non-hibernating animals, which was helpful for the application of clinics and spaceflight.

Simultaneous Observation of Mouse Cortical and Hippocampal Neural Dynamics under Anesthesia through a Cranial Microprism Window.

The fluorescence microscope has been widely used to explore dynamic processes in vivo in mouse brains, with advantages of a large field-of-view and high spatiotemporal resolution. However, owing to background light and tissue scattering, the single-photon wide-field microscope fails to record dynamic neural activities in the deep brain. To achieve simultaneous imaging of deep-brain regions and the superficial cortex, we combined the extended-field-of-view microscopy previously proposed with a novel prism-based cranial window to provide a longitudinal view. As well as a right-angle microprism for imaging above 1 mm, we also designed a new rectangular-trapezoidal microprism cranial window to extend the depth of observation to 1.5 mm and to reduce brain injury. We validated our method with structural imaging of microglia cells in the superficial cortex and deep-brain regions. We also recorded neuronal activity from the mouse brains in awake and anesthesitized states. The results highlight the great potential of our methods for simultaneous dynamic imaging in the superficial and deep layers of mouse brains.

SWCNTs/PEDOT:PSS-Modified Microelectrode Arrays for Dual-Mode Detection of Electrophysiological Signals and Dopamine Concentration in the Striatum under Isoflurane Anesthesia.

Accurate detection of the degree of isoflurane anesthesia during a surgery is important to avoid the risk of overdose isoflurane anesthesia timely. To address this challenge, a four-shank implantable microelectrode array (MEA) was fabricated for the synchronous real-time detection of dual-mode signals [electrophysiological signal and dopamine (DA) concentration] in rat striatum. The SWCNTs/PEDOT:PSS nanocomposites were modified onto the MEAs, which significantly improved the electrical and electrochemical performances of the MEAs. The electrical performance of the modified MEAs with a low impedance (16.20 ± 1.68 kΩ) and a small phase delay (-27.76 ± 0.82°) enabled the MEAs to detect spike firing with a high signal-to-noise ratio (> 3). The electrochemical performance of the modified MEAs with a low oxidation potential (160 mV), a low detection limit (10 nM), high sensitivity (217 pA/µM), and a wide linear range (10 nM-72 µM) met the specific requirements for DA detection in vivo. The anesthetic effect of isoflurane was mediated by inhibiting the spike firing of D2_SPNs (spiny projection neurons expressing the D2-type DA receptor) and the broadband oscillation rhythm of the local field potential (LFP). Therefore, the spike firing rate of D2_SPNs and the power of LFP could reflect the degree of isoflurane anesthesia together. During the isoflurane anesthesia-induced death procedure, we found that electrophysiological activities and DA release were strongly inhibited, and changes in the DA concentration provided more details regarding this procedure. The dual-mode recording MEA provided a detection method for the degree of isoflurane anesthesia and a prediction method for fatal overdose isoflurane anesthesia.

Real-time brain-wide multi-planar microscopy for simultaneous cortex and hippocampus imaging at the cellular resolution in mice.

Interactions between the cerebral cortex and the deep cerebellar nuclei play important roles in cognitive processes. However, conventional microscopes fail to dynamically record cellular structures in distinct brain regions and at different depths, which requires high resolution, large field of view (FOV), and depth of field (DOF). Here we propose a single-photon excited fluorescence microscopy technique that performs simultaneous cortex and hippocampus imaging, enabled by a customized microscope and a chronic optical window. After we implant a glass microwindow above the hippocampus, the surface of the hippocampus is shifted to the superficial plane. We demonstrate that the proposed technique is able to image cellular structures and blood vessel dynamics in the cortex and the hippocampus in in vivo experiments, and is compatible with various mesoscopic systems.

New electrochemical method for programmed death-ligand 1 detection based on a paper-based microfluidic aptasensor.

As programmed death-ligand 1 (PD-L1) is considered a referenced therapeutic biomarker, a rapid and low-cost method to detect PD-L1 in body fluids is necessary. In this work, a paper-based microfluidic aptasensor for label-free electrochemical detection of PD-L1 in liquids was fabricated. The aptasensor integrates a reaction cell and a three-electrode system, and a differential pulse voltammetry electrochemical method was adopted. PD-L1 aptamer with a low equilibrium dissociation constant was used as a biorecognition molecule. To bind the aptamer and assist in the electrochemical measurement, nanocomposites were synthesized and used to modify the working electrode, which was composed of an amine-functionalized single-walled carbon nanotube, new methylene blue and gold nanoparticles. The basic performance of the aptasensor was tested in phosphate-buffered saline: the linear range was between 10 pg mL-1 and 2.5 ng mL-1, and the detection limit was 10 pg mL-1 (signal/noise = 3). Moreover, the aptasensor was used for the detection of serum samples and compared with an enzyme linked immunosorbent assay (ELISA). The results showed that the aptasensor provides a new low-cost, portable and highly sensitive detection method for PD-L1, as an alternative to ELISA.

Cellular-Scale Microelectrode Arrays to Monitor Movement-Related Neuron Activities in the Epileptic Hippocampus of Awake Mice.

OBJECTIVE: Epilepsy affects 50 million people worldwide and its pathogenesis is still unknown. In particular, the movement-related neural activities involving glutamate (Glu) and electrophysiological signals at cellular level remains unclear. METHODS: A cellular-scale implantable microelectrode array (MEA) was fabricated to detect the movement-related neural activities involving Glu concentration and electrophysiological signals. Platinum and reduced graphene oxide nanocomposites were deposited to enhance the surface area. Glu oxidase (Gluox) were coated to effectively recognize Glu molecule. RESULTS: Neural activities in the hippocampus of normal and epileptic mice is different, and the changes are closely connected with movement. Glu concentration and spike firing rate in the epileptic mice were much higher than those in the normal ones. And the neural activities with significant synchronization were detected in the epileptic mice even without seizure occurrence. Meanwhile, the spikes fire more intensively and Glu level became much higher during the movement of the mice compared to the stationary state. CONCLUSION: The existing abnormality of neural activities in the epileptic mice are potential factors to induce a seizure. Movement may impact the neural activities and the duration of seizure. SIGNIFICANCE: The MEA can monitor changes of movement, Glu and neuron discharges synchronously and provides us an effective technology to understand the neuronal disease.

PDMS-Parylene Hybrid, Flexible Micro-ECoG Electrode Array for Spatiotemporal Mapping of Epileptic Electrophysiological Activity from Multicortical Brain Regions.

Epilepsy detection and focus location are urgent issues that need to be solved in epilepsy research. A cortex conformable and fine spatial accuracy electrocorticogram (ECoG) sensor array, especially for real-time detection of multicortical functional regions and delineating epileptic focus remains a challenge. Here, we fabricated a polydimethylsiloxane (PDMS)-parylene hybrid, flexible micro-ECoG electrode array. The multiwalled carbon nanotubes (MWCNTs)/poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) nanocomposite-modified electrode interface significantly improved the sensing performance with low impedance (20.68 ± 6.65 kΩ), stable phase offset, and high sensitivity. The electrophysiological activities of multicortical brain regions (somatosensory cortex, parietal association cortex, and visual cortex) were simultaneously monitored during normal and epileptic statuses. The epileptic ECoG activities spread spatiotemporally from the starting point toward the adjacent cortex. Significant variations of the waveform, power, and frequency band were observed. The ECoG potential (123 ± 23 µV) at normal status was prominently up to 417 ± 87 µV at the spike wave stage. Besides, the power for epileptic activity (11.049 ± 4.513 µW) was 10 times higher than that (1.092 ± 0.369 µW) for normal activity. In addition, the theta frequency band was found to be a characteristic frequency band of epileptic signals. These joint analysis results of multicortical regions indicated that the active micron-scale region on the parietal association cortex was more likely to be the epileptogenic focus. Cortical mapping with high spatial detail provides the accurate delineation of lesions. The flexible micro-ECoG electrode array is a powerful tool for constructing a spatiotemporal map of the cortex. It provides a technical platform for epileptic focus location, biomedical diagnosis, and brain-computer interaction.

Dopamine and Striatal Neuron Firing Respond to Frequency-Dependent DBS Detected by Microelectrode Arrays in the Rat Model of Parkinson's Disease.

(1) Background: Deep brain stimulation (DBS) is considered as an efficient treatment method for alleviating motor symptoms in Parkinson's disease (PD), while different stimulation frequency effects on the specific neuron patterns at the cellular level remain unknown. (2) Methods: In this work, nanocomposites-modified implantable microelectrode arrays (MEAs) were fabricated to synchronously record changes of dopamine (DA) concentration and striatal neuron firing in the striatum during subthalamic nucleus DBS, and different responses of medium spiny projecting neurons (MSNs) and fast spiking interneurons (FSIs) to DBS were analyzed. (3) Results: DA concentration and striatal neuron spike firing rate showed a similar change as DBS frequency changed from 10 to 350 Hz. Note that the increases in DA concentration (3.11 ± 0.67 µM) and neural spike firing rate (15.24 ± 2.71 Hz) were maximal after the stimulation at 100 Hz. The MSNs firing response to DBS was significant, especially at 100 Hz, while the FSIs remained stable after various stimulations. (4) Conclusions: DBS shows the greatest regulatory effect on DA concentration and MSNs firing rate at 100 Hz stimulation. This implantable MEA in the recording of the neurotransmitter and neural spike pattern response to DBS provides a new insight to understand the mechanism of PD at the cellular level.

Paper-based microfluidic aptasensors.

For in-situ disease markers detection, point-of-care (POC) diagnosis has great advantages in speed and cost compared with traditional techniques. The rapid diagnosis, prognosis, and surveillance of diseases can significantly reduce disease-related mortality and trauma. Therefore, increasing attention has been paid to the POC diagnosis devices due to their excellent diagnosis speed and portability. Over the past ten years, paper-based microfluidic aptasensors have emerged as a class of critical POC diagnosis devices and various aptasensors have been proposed to detect various disease markers. However, most aptasensors need further improvement before they can actually enter the market and be widely used. There is thus an urgent need to sort out the key points of preparing the aptasensors and the direction that needs to be invested in. This review summarizes the representative articles in the development of paper-based microfluidic aptasensors. These works can be divided into paper-based optical aptasensors and paper-based electrochemical aptasensors according to their output signals. Significant focus is applied to these works according to the following three parts: (1) The ingenious design of device structure; (2) Application and synthesis of nanomaterial; (3) The detection principle of the proposed aptasensor. This is a detailed and comprehensive review of paper-based microfluidic aptasensors. The accomplishments and shortcomings of the current aptasensors are outlined, the development direction and the future prospective are given. It is hoped that the research in this review can provide a reference for further development of more advanced, more effective paper-based microfluidic aptasensors for POC disease markers diagnosis.

Research on the Characteristics of Action Potentials and Local Field Potentials in Cortex of Parkinson's Disease Mode Monkey.

Parkinson's disease (PD) is characterized by excessively synchronized neural activity. In this paper, we recorded electrophysiological signals in Cortex of normal and PD mode monkey using homemade implantable microelectrode arrays (MEA), and analyzed the characteristics of action potentials (APs) and local field potentials (LFPs). Results showed that, comparing to normal monkey, the spike-firing activity of PD mode monkey could be divided into two stages: the continuous spike-firing stage and the burst spike-firing stage. The continuous spike-firing lasted for about 20s and oscillated at low frequency about 0.03Hz. APs fired in a burst mode between two continuous discharges. In the continuous spike-firing stage, the spike-firing activity was related to the ripple rhythm (100-200Hz) of LFPs with a coherence 0.86, while, in the burst spike-firing stage, it was related to the phase of theta rhythm (4-7 Hz). APs tended to discharge in the valley of theta rhythm (average peak phase is -10°).Clinical Relevance- This article can provide some references for the study of PD neuropathology.

CB1-Antibody Modified Liposomes for Targeted Modulation of Epileptiform Activities Synchronously Detected by Microelectrode Arrays.

Temporal lobe epilepsy (TLE) is a focal, recurrent, and refractory neurological disorder. Therefore, precisely targeted treatments for TLE are greatly needed. We designed anti-CB1 liposomes that can bind to CB1 receptors in the hippocampus to deliver photocaged compounds (ruthenium bipyridine triphenylphosphine γ-aminobutyric acid, RuBi-GABA) in the TLE rats. A 16-channel silicon microelectrode array (MEA) was implanted for simultaneously monitoring electrophysiological signals of neurons. The results showed that anti-CB1 liposomes were larger in size and remained in the hippocampus longer than unmodified liposomes. Following the blue light stimulation, the neural firing rates and the local field potentials of hippocampal neurons were significantly reduced. It is indicated that RuBi-GABA was enriched near hippocampal neurons due to anti-CB1 liposome delivery and photolyzed by optical stimulation, resulting dissociation of GABA to exert inhibitory actions. Furthermore, K-means cluster analysis revealed that the firing rates of interneurons were decreased to a greater extent than those of pyramidal neurons, which may have been a result of the uneven diffusion of RuBi-GABA due to liposomes binding to CB1. In this study, we developed a novel, targeted method to regulate neural electrophysiology in the hippocampus of the TLE rat using antibody-modified nanoliposomes, implantable MEA, and photocaged compounds. This method effectively suppressed hippocampal activities during seizure ictus with high spatiotemporal resolution, which is a crucial exploration of targeted therapy for epilepsy.

Flexible Electrocorticography Electrode Array for Epileptiform Electrical Activity Recording under Glutamate and GABA Modulation on the Primary Somatosensory Cortex of Rats.

Epilepsy is a common neurological disorder. There is still a lack of methods to accurately detect cortical activity and locate lesions. In this work, a flexible electrocorticography (ECoG) electrode array based on polydimethylsiloxane (PDMS)-parylene was fabricated to detect epileptiform activity under glutamate (Glu) and gamma-aminobutyric acid (GABA) modulation on primary somatosensory cortex of rats. The electrode with a thickness of 20 µm has good flexibility to establish reliable contact with the cortex. Fourteen recording sites with a diameter of 60 µm are modified by electroplating platinum black nanoparticles, which effectively improve the performance with lower impedance, obtaining a sensitive sensing interface. The electrode enables real-time capturing changes in neural activity under drug modulation. Under Glu modulation, neuronal populations showed abnormal excitability, manifested as hypsarrhythmia rhythm and continuous or periodic spike wave epileptiform activity, with power increasing significantly. Under GABA modulation, the excitement was inhibited, with amplitude and power reduced to normal. The flexible ECoG electrode array could monitor cortical activity, providing us with an effective tool for further studying epilepsy and locating lesions.

Implanted multichannel microelectrode array for simultaneous electrophysiological signal detection of hippocampal CA1 and DG neurons of simulated microgravity rats.

Microgravity can cause body fluids to accumulate in the brain, resulting in brain damage. There are few studies that focus on the detection of electrophysiological signals in simulated microgravity rats, and the precise mechanisms are unknown. In this study, a new device was established to investigate the influence of microgravity on hippocampal neurons. A 16-channel microelectrode array was fabricated for in vivo multichannel electrophysiological recordings. In these experiments, microelectrode array was inserted into normal, 28-day tail suspension model, and 3-day recovered after modulation rats to record electrophysiological signals in the CA1 and DG regions of the hippocampus. Through analysis of electrophysiological signals, we obtained the following results: (1) spike signals of model rats sporadically showed brief periods of suspension involving most of the recorded neurons, which corresponded to slow and smooth peaks in local field potentials. For model rats, the firing rate was reduced, and the power in the frequency spectrum was concentrated in the slow frequency band (0-1 Hz); (2) after the detected hippocampal cells divided into pyramidal cells and interneurons, the spike duration of pyramidal cells showed remarkable latency, and their average firing rates showed a more significant decrease compared to interneurons. These results demonstrate that the hippocampal neurons were impaired after modulation in the cellular dimension, and pyramidal cells were more susceptible than interneurons.