A mouse sleep fragmentation research system based on electrophysiological electrochemical synchronous recording
By designing a mouse sleep fragmentation research system that simultaneously records electrophysiological and electrochemical signals, we have achieved mouse sleep fragmentation modeling and simultaneous recording of electrophysiological and electrochemical signals. This solves the problem that existing technologies cannot simultaneously monitor changes in brain neurochemicals, provides a complete experimental system, and reveals the impact of sleep fragmentation on the brain.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-03-26
- Publication Date
- 2026-06-23
AI Technical Summary
Existing small animal sleep research systems cannot achieve simultaneous recording of electrophysiological and electrochemical signals during the modeling process, lack the ability to detect brain waves and neurochemical substances in situ, and make it difficult to assess the neurochemical processes of sleep fragmentation and the degree of modeling.
A mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording was designed, including a sleep fragmentation modeling module, a signal measurement and acquisition control module, and an analysis module. A flexible interference rod driven by a programmable stepper motor is used to interfere with sleep. An electrophysiological detection unit and an electrochemical detection unit are integrated to realize the simultaneous acquisition and analysis of electrophysiological and electrochemical signals.
This study enabled the simultaneous recording of electrophysiological and electrochemical signals in mouse sleep fragmentation modeling, providing the ability to detect neurochemicals in the brain in situ and quantitatively assess the impact of sleep fragmentation, thus providing a complete experimental system for the study of the neurochemical mechanisms of sleep fragmentation.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of animal experimental equipment technology, specifically relating to a mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording. Background Technology
[0002] Sleep is a key physiological process for maintaining homeostasis and higher cognitive functions. Sleep fragmentation, a common sleep disorder, is characterized by an abnormal increase in the number of micro-awakenings and a significant decrease in the proportion of deep sleep, even when the total sleep duration remains relatively constant. It can be caused by various factors such as obstructive sleep apnea, chronic pain, and aging, and can lead to physiological and psychological problems such as memory loss, cognitive decline, and metabolic disorders. Therefore, it is of great significance to study the impact of chronic sleep fragmentation on the homeostasis of neurochemical substances in organisms.
[0003] Current research on sleep fragmentation is moving from macroscopic clinical phenomena to microscopic molecular mechanisms, focusing on the release and regulation of neurotransmitters and neuromodulators and their association with diseases. However, in vivo in situ detection of changes in neurochemical substances in complex brain tissue remains a pressing scientific problem in this field. Existing small animal sleep research systems have the following problems: they primarily function as sleep disturbance models, lacking the ability to simultaneously record electrophysiological and electrochemical signals during the modeling process, thus failing to reflect the neurophysiological status of the rat brain; they lack the ability to record key physiological indicators such as electroencephalograms and neurochemical substances in situ, making it difficult to support research on the neurochemical processes of sleep fragmentation; and they cannot assess the degree of modeling and the neurochemical impact.
[0004] Therefore, how to provide a mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention
[0005] Based on the above analysis, this invention aims to provide a mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording, addressing the problems of existing small animal sleep research systems: primarily focused on sleep disturbance modeling, lacking simultaneous recording of electrophysiological and electrochemical signals during modeling, thus failing to reflect the neurophysiological state of the mouse brain; lacking in-situ recording capabilities for key physiological indicators such as EEG and neurochemical substances, making it difficult to support research on brain neurochemical processes in sleep fragmentation; and exhibiting low accuracy in assessing the degree of modeling and neurochemical aspects.
[0006] The objective of this invention is mainly achieved through the following technical solutions: The first aspect of the present invention provides a mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording, which includes a sleep fragmentation modeling module, a signal measurement and acquisition control module, and an analysis module; The sleep fragmentation modeling module provides a sleep space for mice and disrupts their sleep at predetermined intervals; the signal measurement and acquisition control module measures and acquires the electrophysiological and electrochemical signals of the disturbed mice; and the analysis module generates key sleep parameters based on the electrophysiological and electrochemical signals to evaluate sleep quality and modeling effectiveness.
[0007] Furthermore, the sleep fragmentation modeling module includes a sleep chamber and an interference device, wherein the interference device is disposed inside the sleep chamber; the mouse is disposed inside the sleep chamber, and the interference device is used to interfere with the mouse's sleep.
[0008] Furthermore, the sleeping box includes a food box, a water box, and a lighting system.
[0009] Furthermore, the sleep box includes a sleep disturbance module, which is disposed on the lower end face inside the sleep box.
[0010] Furthermore, the signal measurement and acquisition control module includes an electrophysiological detection unit and an electrochemical detection unit.
[0011] Furthermore, the electrophysiological detection unit includes electrophysiological detection electrodes, which include: a parietal lobe electrode implanted in the parietal lobe of a mouse, an EEG electrode implanted in the frontal lobe brain region, a ground electrode implanted in the cerebellum, and an electromyographic electrode implanted in the neck and back muscles of a mouse.
[0012] Furthermore, the electrochemical detection unit includes an electrochemical detection electrode, which includes a carbon fiber microelectrode and a micro all-solid-state ion-selective electrode.
[0013] Furthermore, the carbon fiber microelectrode and the miniature all-solid-state ion-selective electrode were implanted into the nucleus accumbens region of the mouse brain.
[0014] A second aspect of the present invention provides a method for establishing a mouse sleep fragmentation model and conducting electrophysiological and electrochemical assessment using the above-described research system, comprising the following steps: Step 1: Animal pretreatment. Electrophysiological and electrochemical detection electrodes are placed at the animal's response sites. Step 2: Sleep fragmentation modeling. Using a light system and a sleep disturbance module, sleep is disturbed in mice to construct an animal sleep fragmentation model, and electrophysiological and electrochemical signals are collected simultaneously. Step 3: Data processing and analysis. Electrophysiological signals are used to identify the sleep state of the animals, and the electrochemical signals are used to evaluate the sleep quality and modeling effect of the animals.
[0015] Furthermore, the sleep states include wakefulness, REM sleep, and non-REM sleep.
[0016] Compared with the prior art, the present invention can achieve at least one of the following beneficial effects: 1. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording described in this invention realizes the integrated design of mouse sleep fragmentation modeling, electrophysiological signal recording, and electrochemical signal detection. It applies the in vivo in situ electrochemical detection method to the study of animal models of sleep fragmentation, solves the technical problem that existing modeling devices cannot simultaneously monitor changes in brain neurochemical substances, and provides a complete experimental system for the study of the neurochemical mechanisms of sleep fragmentation.
[0017] 2. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording described in this invention uses a flexible interference rod driven by a programmable stepper motor in the sleep fragmentation modeling module to achieve programmed sleep disturbance once every 2 minutes, simulating the sleep fragmentation frequency characteristics of human patients with severe obstructive sleep apnea. Furthermore, the design of the silent motor and sound insulation cotton minimizes the impact of environmental stimuli on the experimental results, making the modeling results more consistent with clinical practice.
[0018] 3. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording described in this invention employs a signal acquisition strategy that alternates between voltammetric scanning and potential recording. This effectively reduces the interference of the scanning voltage of the rapid scanning cyclic voltammetry method on electrophysiological and ion potential signals, ensuring the accuracy of electrophysiological and electrochemical signal acquisition and achieving continuous synchronous recording of the two signals.
[0019] 4. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording described in this invention integrates carbon fiber microelectrodes and miniature all-solid-state ion-selective electrodes in its electrochemical detection electrode, simultaneously detecting neurochemical molecules such as dopamine and ascorbic acid in the brain, as well as Ca²⁺. + K + H + The plasma concentration changes, small electrode size, and high spatiotemporal resolution allow for simultaneous implantation with EEG / EMG electrodes, enabling in-situ real-time detection of multiple indicators.
[0020] 5. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording described in this invention can quantitatively assess the degree of sleep fragmentation modeling, reveal the impact of sleep fragmentation on neurotransmitters and ion homeostasis in key brain regions such as the nucleus accumbens, elucidate the neurochemical basis of physiological and cognitive dysfunction caused by sleep fragmentation at the cellular and molecular level, provide a scientific basis for the improvement and treatment strategies of sleep fragmentation, and has important clinical translational value. Attached Figure Description
[0021] Figure 1 This is a schematic diagram of the overall structure of the mouse sleep fragmentation study system with simultaneous electrophysiological and electrochemical recording, as shown in Example 1. Figure 2 This is a graph showing the electrophysiological recording results and sleep state parameter analysis of the sleep fragmentation modeling process in Example 2; Figure 3 This is a bar chart showing the average duration of NREM sleep, REM sleep, and wakefulness in mice of the control and model groups within 24 hours in Example 2. Figure 4 This is a bar chart showing the number of NREM sleep-wake cycles within 24 hours in Example 2; Figure 5 This is a diagram showing the results of neurochemical detection in the nucleus accumbens region of the mouse brain in Example 3.
[0022] Figure label: 1-Sleep fragmentation modeling module, 11-Sleep box, 111-Water box, 112-Food box, 113-Camera, 114-Lighting system, 115-Sound insulation cotton, 116-Exhaust fan, 12-Interference rod, 13-Sleep interference module, 2-Signal measurement and acquisition control module, 21-Sleep fragmentation modeling control interface, 22-Electrochemical and electrophysiological signal acquisition interface, 23-Video data acquisition interface. Detailed Implementation
[0023] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. The described embodiments are only used to explain the present invention and are not intended to limit the scope of protection of the present invention.
[0024] A mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording, such as Figure 1 As shown, it includes a sleep fragmentation modeling module 1, a signal measurement and acquisition control module 2, and an analysis module; The sleep fragmentation modeling module 1 provides a sleep space for mice and disrupts their sleep at predetermined intervals; the signal measurement and acquisition control module 2 is used to measure and acquire the electrophysiological and electrochemical signals of the disturbed mice; and the analysis module is used to generate key sleep parameters based on the electrophysiological and electrochemical signals to evaluate sleep quality and modeling effect.
[0025] The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording described in this invention realizes the integrated design of mouse sleep fragmentation modeling, electrophysiological signal recording, and electrochemical signal detection. It applies the in vivo in situ electrochemical detection method to the study of animal models of sleep fragmentation, solves the technical problem that existing modeling devices cannot simultaneously monitor changes in brain neurochemical substances, and provides a complete experimental system for the study of the neurochemical mechanisms of sleep fragmentation.
[0026] Furthermore, the sleep fragmentation modeling module 1 includes a sleep box 11 and an interference device, the interference device being disposed inside the sleep box 11; the mouse is disposed inside the sleep box 11, and the interference device is used to interfere with the mouse's sleep.
[0027] The sleeping box 11 includes a food box 112, a water box 111, and a lighting system 114. The inner wall of the sleeping box is lined with sound-absorbing cotton. The food box 112 and water box 111 are located on the side wall of the sleeping box 11. The lighting system 114 is located on the upper surface inside the sleeping box 11 and is used to simulate circadian rhythms. The interference device includes a programmable stepper motor and a rotatable interference rod 12 driven by it. The programmable stepper motor is configured to drive the interference rod 12 to move once every 2 minutes, with each movement lasting 20 seconds. The movement consists of one clockwise rotation followed by one counterclockwise rotation. The rotatable interference rod 12 is made of flexible material.
[0028] The lighting system 114 is configured to simulate a 12h / 12h light-dark cycle. During the light cycle, the programmable stepper motor is activated to induce sleep disturbance, and during the dark period, the programmable stepper motor stops working.
[0029] The sleep chamber 11 is equipped with an exhaust fan and a video acquisition device, which is a camera 113 located inside the sleep chamber 11, used to collect behavioral videos of mice.
[0030] Furthermore, the signal measurement and acquisition control module 2 includes an electrophysiological signal recording device, which includes an electrophysiological detection unit, an electrochemical detection unit, and a signal amplifier.
[0031] Furthermore, the signal measurement and acquisition control module 2 includes a sleep fragmentation modeling control interface 21, an electrochemical and electrophysiological signal acquisition interface 22, and a video data acquisition interface 23.
[0032] The electrophysiological detection unit includes electrophysiological detection electrodes, which include a parietal lobe electrode implanted in the parietal lobe of a mouse, an EEG electrode implanted in the frontal lobe, a ground electrode implanted in the cerebellum, and an electromyographic electrode implanted in the neck and back muscles of a mouse; the parietal lobe electrode, the EEG electrode, the ground electrode, and the electromyographic electrode are integrated into the same female interface.
[0033] The electrochemical detection unit includes an electrochemical detection electrode and a voltammetry-potential detection unit. The electrochemical detection electrode is integrated with an EEG / EMG integrated electrode to the same interface. The electrochemical detection electrode includes a carbon fiber microelectrode and a miniature all-solid-state ion-selective electrode. The voltammetry-potential detection unit is configured to detect electrochemically active neurochemical molecules in the brain using a rapid scanning cyclic voltammetry method via the carbon fiber microelectrode, and to detect the ion concentration in the brain via the miniature all-solid-state ion-selective electrode.
[0034] The electrochemically active neurochemical molecules include dopamine and ascorbic acid, and the intracranial ions include Ca²⁺. + K + H + One or more of them.
[0035] The system employs a signal acquisition strategy that alternates between voltammetric scanning and potential recording: the scanning process of rapid scanning cyclic voltammetry is performed alternately with the electrophysiological signal recording process, and the potential signal recording process of ion-selective electrodes is acquired synchronously with the electrophysiological signal recording process; wherein the scanning frequency of rapid scanning cyclic voltammetry is 5~50Hz, and the scan rate is 400~1000V / s.
[0036] The electrophysiological detection electrode and the electrochemical detection electrode are fixed to the mouse skull with dental cement, and the electrochemical detection electrode is implanted in the nucleus accumbens region of the mouse brain.
[0037] A second aspect of the present invention also provides a method for establishing a mouse sleep fragmentation model and conducting electrophysiological and electrochemical assessment using the above-described research system, comprising the following steps: Step 1: Animal pretreatment. Electrophysiological and electrochemical detection electrodes are placed at the animal's response sites. Step 2: Sleep fragmentation modeling. Using the light system 114 and the sleep disturbance module 13, sleep disturbance is performed on mice to construct an animal sleep fragmentation model and collect electrophysiological and electrochemical signals. Step 3: Data processing and analysis. Electrophysiological signals are used to identify the sleep state of the animals, and the electrochemical signals are used to evaluate the sleep quality and modeling effect of the animals.
[0038] Furthermore, the sleep states include wakefulness, REM sleep, and non-REM sleep.
[0039] Furthermore, in step two, the electrophysiological and electrochemical signals are acquired according to the following signal acquisition strategy: Step 1: Preset the basic cycle of the voltammetric scan. The scan frequency is 5~50Hz, that is, the detection cycle for completing one fast scan cyclic voltammetry is 200ms (5Hz) to 20ms (50Hz). This cycle is the smallest time unit of timing planning, denoted as cycle T.
[0040] Step 2: Within the basic cycle, the first signal and the second signal are acquired sequentially to complete the signal acquisition for one basic cycle; the first signal is an electrochemical voltammetric signal, and the second signal includes electrophysiological signals and ion potential acquisition.
[0041] Step 3: Repeat the signal acquisition steps of Step 2 in the next cycle.
[0042] Furthermore, the signal measurement and acquisition control module 2 is electrically connected to the sleep fragmentation modeling module 1 and the analysis module, respectively, to realize the synchronous regulation of modeling program control, electrophysiological signal acquisition, video data acquisition and electrochemical signal detection.
[0043] Furthermore, the ion site acquisition employs a miniature all-solid-state ion-selective electrode, which is a modified electrode based on a carbon fiber electrode, including a pH electrode and a Ca²⁺ electrode. + Selective electrode, K + One or more of the selective electrodes.
[0044] Preferably, the miniature all-solid-state ion-selective electrode is Ca²⁺. + Selective electrode, the Ca² + The method for preparing the selective electrode is as follows: S1. Preparation of carbon fiber electrodes; S2. Electrochemical acid pretreatment activation of carbon fiber electrodes; S3. The carbon fiber electrode tip was immersed in a 2 mg / mL three-shell hollow carbon sphere dispersion for rolling modification. After drying at room temperature, the process was repeated several times to obtain a carbon fiber electrode modified with three-shell hollow carbon spheres. S4. Immerse the modified carbon fiber electrode in a droplet of calcium ion-sensitive membrane and roll it to modify it until the electrode tip is completely covered, thus obtaining the initial Ca²⁺. + Selective electrode; S5, Prepare the initial Ca² + The selective electrode was activated by immersion in 0.1 M CaCl2 solution for at least 24 hours to obtain the finished Ca²⁺ product. + Selective electrode.
[0045] The following sections verify the integrity and practicality of the research system by constructing a mouse sleep fragmentation model, performing electrophysiological analysis of sleep states during the modeling process, and conducting in-situ detection of neurochemical molecules and ions in the brain after modeling. All embodiments used adult C57BL / 6J mice as experimental subjects, strictly followed animal ethics standards, and adopted a 12h / 12h light-dark cycle for feeding (lights on at 8 am and off at 8 pm). Quantitative data are expressed as mean ± standard deviation (Mean ± SEM), and paired t-tests were used to analyze differences (P < 0.05 was considered statistically significant). The detailed schemes, objectives, and effects of each embodiment are as follows.
[0046] Example 1: Construction of a mouse sleep fragmentation model 1.1 Purpose of Implementation: To construct a mouse sleep fragmentation model simulating human severe obstructive sleep apnea (OSA) patients through programmed mechanical perturbation, providing a standardized experimental model for subsequent sleep state analysis and neurochemical detection in the brain, and at the same time verifying the programmed interference function of sleep fragmentation modeling module 1.
[0047] 1.2 Implementation Plan Modeling equipment: The sleep fragmentation modeling module 1 designed according to this invention is as follows Figure 1 As shown, the sleep fragmentation modeling module 1 includes a sleep chamber 11 and an interference device. The interference device includes a movable column, an interference rod 12, and a programmable silent stepper motor. The interference rod 12 is mounted on the movable column. The programmable silent stepper motor drives the movable column to rotate, which in turn drives the interference rod 12 to rotate, thus interfering with the mouse's sleep. The sleep chamber 11 has a water box 111 and a food box 112 on its side wall. The sleep chamber 11 is equipped with sound insulation cotton and an exhaust fan 116, among other auxiliary devices.
[0048] Interference Program: A programmable stepper motor was used to disturb mice every 2 minutes during their sleep period (during light exposure), with each disturbance lasting 20 seconds. During interference, the motor drove the interference rod 12 to rotate clockwise once and then counterclockwise once, gently disturbing the mice's sleep through physical contact. During the mice's awake period (during darkness), the motor remained stationary, with no intervention. Mice are nocturnal animals, so during light exposure, which corresponds to the mice's sleep period, the programmable stepper motor was activated to disrupt their sleep; during darkness, i.e., during wakefulness, the motor and the crossbar remained stationary, with no intervention. This was continued for at least 14 consecutive days to establish a mouse model of fragmented sleep.
[0049] Experimental grouping: A sleep fragmentation model group and a control group were set up. The control group mice were placed in the same environment without sleep disturbance. Four mice were selected, and their normal sleep state was recorded as their own control. Then, the sleep fragmentation model was established for at least 14 consecutive days. Sleep data were recorded again after the model was established.
[0050] Electrode Implantation: Mice were anesthetized using a gas anesthesia machine. Partial hair was shaved from the mouse skull, and the mouse head was fixed using a stereotaxic instrument. The skull skin was cut open to expose the skull, and the surface mucosa was cleaned with 3% hydrogen peroxide to remove corrosion, followed by cleaning with physiological saline. Based on a stereotaxic map of the mouse brain, holes were drilled in the tops of the frontal bone (AP = 1.0 mm, ML = ±1.3 mm), parietal bone (AP = -3 mm, ML = ±2.5 mm), and cerebellum (AP = -5 mm, ML = ±1.0 mm), penetrating the skull without damaging brain tissue. Screws with welded wires were screwed into the frontal and parietal bones to serve as EEG electrodes for acquiring cortical EEG signals, while a screw with welded wires was screwed into the top of the cerebellum as a ground wire and reference. Two stainless steel wires were implanted as electromyography (EMG) electrodes in the neck and back muscles of the mice to acquire EMG signals. All the above wires were integrated together for connection to a sleep monitoring instrument. Simultaneously, referring to brain atlases, a 2mm × 2mm window was opened above the target brain region for electrochemical detection using a cranial drill, and electrodes or ion-selective electrodes for rapid scanning cyclic voltammetry were implanted into the target brain region. All electrodes were fixed with dental cement. After the mice had recovered for at least 7 days, they were placed in a sleep fragmentation instrument for at least 3 days to acclimatize.
[0051] Implementation effect A standardized and reproducible model of fragmented sleep in mice was established. Mechanical disturbances were used as gentle physical stimuli, which minimized the additional stress on mice and reduced model bias. The programming function of the modeling equipment matches the frequency of sleep fragmentation in human patients with severe OSA, and the model has high clinical relevance.
[0052] Example 2: Electrophysiological analysis of mouse sleep states during sleep fragmentation modeling 2.1 Implementation Objectives Using the electrophysiological detection unit and video acquisition equipment in the signal measurement and acquisition control module 2, the electroencephalogram (EEG), electromyography (EMG), and behavioral signals of mice before and after modeling were recorded to quantitatively evaluate the effect of sleep fragmentation modeling and verify the system's ability to identify and analyze mouse sleep states in real time.
[0053] 2.2 Implementation Plan Signal acquisition: Cortical electroencephalogram (EEG) signals and neck and back muscle electroencephalogram (EEG) signals of mice in a free-moving state were acquired by electrophysiological detection electrodes. Behavioral videos were simultaneously acquired by camera 113. The electrophysiological signals were transmitted to a high-performance acquisition system using a signal amplifier. Monitoring was conducted throughout the process before, during, and after modeling.
[0054] Sleep state recognition: such as Figure 2As shown, based on the typical waveform characteristics of EEG and EMG signals, three states of sleep in mice are identified: wake, REM sleep, and non-REM sleep. Awake phase: EEG desynchronization and low amplitude, EMG high amplitude waveform with explosive discharge; REM phase: EEG is dominated by theta waves (4~10Hz), and EMG amplitude is low and stable; NREM phase: EEG is dominated by delta waves (0.5~4Hz) with high amplitude, while EMG does not have high-amplitude discharge.
[0055] Quantitative evaluation indicators: Statistically analyze and compare the core sleep parameters of mice before and after modeling: ① Duration and proportion of the three sleep / wake states; ② Frequency of micro-arousals and duration of each REM and NREM sleep interval; ③ Number of transitions between the three states (with a focus on the number of transitions between NREM and Wake).
[0056] 2.3 Implementation Results Real-time monitoring of the modeling process: Electrophysiological data captured during the 1-hour modeling process showed that the root mean square of electromyography in mice increased significantly every 2 minutes, and NREM sleep (mainly delta waves) was significantly interrupted, which was completely consistent with the interference frequency set in the program, verifying the accuracy of the modeling module. Compensatory behavior was observed in the early stages of the model: On the first day of the disturbance, the mice's daytime sleep time decreased significantly and their nighttime sleep time increased, reflecting the mice's initial physiological compensation for sleep disturbance. Verification of modeling success: After 14 consecutive days of modeling, the 24-hour sleep-wake rhythm of mice was significantly disrupted, such as... Figure 3 The average duration of each NREM was significantly shortened, as shown in the figure. Figure 4 The number of transitions between NREM and Wake shown increased significantly, demonstrating a significant improvement in the fragmentation of mouse sleep, indicating successful model construction. The functionality and effectiveness of the sleep state analysis module were verified: it can identify three sleep / wake states in mice, realize the quantitative statistics of sleep parameters, and provide an objective and quantifiable evaluation basis for the modeling effect.
[0057] Example 3: In-situ detection of neurochemical molecules and ions in the brain of a mouse model of sleep fragmentation 3.1 Implementation Objectives Using a real-time brain chemical signal measurement module, combined with rapid scanning cyclic voltammetry and a miniature all-solid-state ion-selective electrode, changes in the concentrations of electrochemically active neurochemical molecules (dopamine, ascorbic acid) and ions in the brain of mice after sleep fragmentation modeling were detected in situ and quantitatively. This verified the core function of the system in synchronous recording of electrophysiological and electrochemical signals and revealed the impact of sleep fragmentation on neurochemical homeostasis in the brain.
[0058] 3.2 Implementation Plan This embodiment is divided into two parts: neurochemical molecular detection and ion concentration detection. Both parts employ a signal acquisition strategy of alternating voltammetric scanning and potential recording to ensure that electrophysiological and electrochemical signals are recorded synchronously without interference. 3.2.1 Detection of dopamine and ascorbic acid (rapid scanning cyclic voltammetry) Electrodes and Methods: Carbon fiber microelectrodes were implanted into the nucleus accumbens (target brain region) of mice. Rapid scanning cyclic voltammetry was performed using a scanning frequency of 5–50 Hz and a scan rate of 400–1000 V / s. High concentrations of K+ were used for detection. + Under stimulation (2 μL / min, 30 s), the voltammetric response signals of dopamine and ascorbic acid in the brain were collected; Data Analysis: (1) Acquire raw voltammetric data using the fast scan cyclic voltammetry method; (2) Preprocess the data and construct the dataset; (3) Use a trained generative deep learning model to perform data analysis on the dataset, and separate the ion change signal and the redox voltammetric response of electrochemically active molecules; (4) Quantitative analysis of the redox voltammetric response is performed using a trained deep learning regression model, such as... Figure 5 The curves showing the changes in ascorbic acid, dopamine, and ion concentrations are shown. Substituting the ion change signals into a linear equation yields the ion concentration change results. Figure 5 Curve a represents the quantitative results of ascorbic acid. Figure 5 Curve b represents the quantitative results of dopamine. Figure 5 Curve c in the middle represents the quantitative result of ion concentration.
[0059] 3.2.2 Ion Concentration Detection (Miniature All-Solid-State Ion Selective Electrode Method) Electrode fabrication: Using carbon fiber electrodes as a substrate, the electrodes underwent electrochemical acid pretreatment, 3S-HCNs modification, and Ca... ²+ Sensitive membrane (Ca ²+ -ISM) coating to prepare micro Ca ²+ Selective electrodes should be activated in 0.1 M CaCl2 solution for at least 24 hours before use. Signal acquisition: Ca ²+ Selective electrodes were implanted into the mouse brain simultaneously with EEG / EMG electrodes. Ion potential signals were recorded using the potentiometric method. These signals were acquired synchronously with electrophysiological signals without external voltage interference.
[0060] Signal acquisition strategy: The scanning process of rapid scanning cyclic voltammetry is alternated with the electrophysiological signal recording process (to avoid electrical interference from the scanning voltage). The potential recording of ion-selective electrodes is synchronously acquired with the electrophysiological signals, ultimately achieving continuous and synchronous recording of EEG, EMG, neurochemical molecules, and ion signals during the sleep cycle.
[0061] Implementation effect In situ quantitative detection of neurochemical molecules in the brain was achieved: in K + Under stimulation, the dynamic changes in the concentrations of dopamine and ascorbic acid in the nucleus accumbens of mice were successfully detected. The separation and quantification of signals were achieved through a deep learning model, which solved the problem of interference in the detection of neurochemical molecules in complex brain tissue. Simultaneous monitoring of intracranial ion concentration was achieved: the fabricated miniature all-solid-state ion-selective electrode is small in size and has high spatiotemporal resolution, allowing for synchronous implantation with EEG / EMG electrodes, thus enabling Ca2+ monitoring. ²+ The simultaneous recording of plasma concentration and sleep state provides data for studying the relationship between sleep fragmentation and ion homeostasis; The system's core innovation was verified: through integrated electrode design and single-acquisition-end timing control, interference-free synchronous recording of electrophysiological and electrochemical signals was successfully achieved, solving the technical challenge that existing technologies cannot simultaneously monitor sleep state and neurochemical changes in the brain. The study revealed the neurochemical effects of sleep fragmentation: In vivo in situ electrochemical detection was applied to a sleep fragmentation model, and it was found that sleep fragmentation can lead to changes in the release of dopamine, ascorbic acid, and ionic strength in the brain. This provides experimental evidence for further investigation into the neurochemical mechanisms by which sleep fragmentation causes cognitive dysfunction and metabolic disorders.
[0062] The experimental results of this embodiment demonstrate that the system can simulate the sleep fragmentation characteristics of human OSA patients, and achieve in-situ, real-time, and interference-free synchronous recording of sleep state and neurochemical signals in the brain. It provides a standardized experimental system for in-depth research on the neurochemical regulation mechanism of sleep fragmentation and for finding the key points of its physiological harm, and also provides a scientific basis for the improvement or treatment strategies of sleep fragmentation.
[0063] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.
Claims
1. A mouse sleep fragmentation study system based on electrophysiological electrochemical synchronous recording, characterized in that, It includes a sleep fragmentation modeling module (1), a signal measurement and acquisition control module (2), and an analysis module; The sleep fragmentation modeling module (1) provides a sleep space for mice and interferes with their sleep at predetermined intervals; the signal measurement and acquisition control module (2) is used to measure and acquire the electrophysiological and electrochemical signals of the disturbed mice; the analysis module is used to generate key sleep parameters based on the electrophysiological and electrochemical signals to evaluate sleep quality and modeling effect.
2. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording according to claim 1, characterized in that, The sleep fragmentation modeling module (1) includes a sleep box (11) and an interference device. The interference device is set inside the sleep box (11). The mouse is set inside the sleep box (11). The interference device is used to interfere with the mouse's sleep.
3. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording according to claim 2, characterized in that, The sleeping box (11) includes a food box (112), a water box (111), and a lighting system (114).
4. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording according to claim 2, characterized in that, The sleep fragmentation modeling module (1) also includes a sleep disturbance module (13), which is disposed on the lower end face inside the sleep box (11).
5. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording according to claim 1, characterized in that, The signal measurement and acquisition control module (2) includes an electrophysiological detection unit and an electrochemical detection unit.
6. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording according to claim 5, characterized in that, The electrophysiological detection unit includes electrophysiological detection electrodes; The electrophysiological detection electrodes include: a parietal lobe electrode implanted in the parietal lobe of a mouse, an EEG electrode implanted in the frontal lobe, a ground electrode implanted in the cerebellum, and an electromyographic electrode implanted in the neck and back muscles of a mouse.
7. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording according to claim 5, characterized in that, The electrochemical detection unit includes an electrochemical detection electrode, which includes a carbon fiber microelectrode and a micro all-solid-state ion-selective electrode.
8. The mouse sleep fragmentation research system based on simultaneous electrophysiological and electrochemical recording according to claim 7, characterized in that, The carbon fiber microelectrode and the miniature all-solid-state ion-selective electrode were implanted into the nucleus accumbens region of the mouse brain.
9. A method for establishing a mouse sleep fragmentation model and conducting electrophysiological and electrochemical assessment using the research system described in any one of claims 1-8, characterized in that, Includes the following steps: Step 1: Animal pretreatment. Electrophysiological and electrochemical detection electrodes are placed at the animal's response sites. Step 2: Sleep fragmentation modeling. The sleep of mice was disturbed by the light system (114) and the sleep disturbance module (13) to construct an animal sleep fragmentation model. Electrophysiological and electrochemical signals were collected simultaneously. Step 3: Data processing and analysis. Electrophysiological signals are used to identify the sleep state of the animals, and the electrochemical signals are used to evaluate the sleep quality and modeling effect of the animals.
10. The method according to claim 9, characterized in that, The sleep states include wakefulness, REM sleep, and non-REM sleep.