Realization method and acquisition system of neuroscience-oriented minimally invasive sensor

Abstract

The invention discloses a neuroscience-oriented minimally invasive sensor implementation method and a neuroscience-oriented minimally invasive sensor acquisition system. The method comprises the following steps: preparing a planar microneedle, preparing a neural action potential sensing module on the front surface of the planar microneedle, and preparing a biochemical signal sensing module on the back surface of the planar microneedle; the biochemical signal sensing module comprises a dopamine electrode and an acetylcholine electrode; and overlapping and integrating a plurality of planar microneedles to obtain the minimally invasive sensor. The minimally invasive sensor prepared by the invention is a multifunctional microneedle array sensor, and can synchronously record electroneurographic signals and biochemical signals with high temporal-spatial resolution; and through superposition and integration of the planar microneedle array, the process difficulty of micromachining of the three-dimensional microneedle array is relieved, the preparation process of the minimally invasive sensor is reduced, and the collection effect of the minimally invasive sensor is improved. The method can be widely applied to the technical field of biomedical engineering micro devices.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical engineering micro-devices, in particular to a method for implementing a minimally invasive sensor for neuroscience and an acquisition system. Background Art

[0002] The field of biomedical engineering micro-devices is a research field based on the high cross of materials - micro-devices - biology. Many remarkable achievements have been made in this field in the forefront directions such as cell sensors, microfluidic devices, molecular detection chips, wearable sensors, etc. Bioinformation sensing provides an important technical means for revealing the mysteries of life and guiding disease treatment. People have always been eager to understand nearly a hundred billion neurons in the brain and find a series of new methods for treating neurological diseases such as Parkinson's syndrome, epilepsy, Alzheimer's disease, etc. The sensing detection of physiological indicators of nerves, including nerve electrical signals, biochemical signals, gene information, etc., is a key to unlocking the mystery of nerves and depends on being carried out layer by layer from different levels such as isolated cells, isolated tissues, and living animals. The nerve microelectrode technology is a crucial tool for obtaining nerve electrical signals.

[0003] In the related art, at the in vivo level, in addition to the action potential of nerve cells, the biochemical signals of nerve cells, such as dopamine, acetylcholine, glutamate, etc., also closely affect and mark the processes of various neurological diseases. The sensing of these biochemical signals depends on the electrochemical three-electrode system and specific enzyme modification to achieve specific sensing. Due to the complex process of constructing and modifying precise electrochemical electrodes on the surface of three-dimensional micro-needles, the sensing technology for high spatio-temporal resolution acquisition of biochemical signals in the brain has not been realized. Existing implanted brain electrodes mainly have the function of local electrical signal acquisition, but the long-term acquisition of intracerebral biochemical information has not been realized. Due to the lack of a sensing electrode technology for high-resolution synchronous recording of nerve electrical signals and biochemical signals, the spatio-temporal coupling relationship between brain nerve electrical signals and biochemical signals has not been clarified. Summary of the Invention

[0004] The purpose of the present invention is to solve at least to a certain extent one of the technical problems existing in the prior art.

[0005] To this end, the purpose of the present invention is to provide an efficient method for implementing a minimally invasive sensor for neuroscience and an acquisition system.

[0006] To achieve the above technical objectives, on the one hand, an embodiment of the present invention provides a method for implementing a minimally invasive sensor for neuroscience, including the following steps: preparing planar microneedles, preparing a neural action potential sensing module on the front side of the planar microneedles, and preparing a biochemical signal sensing module on the back side of the planar microneedles; the biochemical signal sensing module includes a dopamine electrode and an acetylcholine electrode; stacking and integrating a plurality of the planar microneedles to obtain a minimally invasive sensor. The minimally invasive sensor prepared in this application is a multifunctional microneedle array sensor that can synchronously record neural electrical signals and biochemical signals with high spatio-temporal resolution; and by stacking and integrating the planar microneedle array, the process difficulties of three-dimensional microneedle array microfabrication are alleviated, which is beneficial to reducing the preparation process of the minimally invasive sensor and improving the acquisition effect of the minimally invasive sensor.

[0007] In some embodiments, for the method for implementing a minimally invasive sensor for neuroscience according to an embodiment of the present invention, the steps of preparing planar microneedles, preparing a neural action potential sensing module on the front side of the planar microneedles, and preparing a biochemical signal sensing module on the back side of the planar microneedles include:

[0008] Preparing planar microneedles by laser micro-cutting a polyimide substrate;

[0009] Preparing multi-channel platinum electrode micro-modules and micro-lines on the front and back sides of the planar microneedles by photolithography;

[0010] Spin-coating a polyimide thin layer on the micro-lines on the front side to expose the front-end microelectrodes, obtaining a neural action potential sensing module;

[0011] Covering the microelectrodes on the back side with a mask plate, and respectively modifying acetylcholinesterase and Ag / AgCl to obtain a biochemical signal sensing module.

[0012] In some embodiments, in one embodiment of the present invention, the method further includes:

[0013] Performing oxygen plasma surface activation on the polyimide surface of the minimally invasive sensor and coupling it with a zwitterionic polymer to form a nanoscale surface coating.

[0014] In some embodiments, in one embodiment of the present invention, the method further includes:

[0015] Optimizing the coupling density of the surface coating of the minimally invasive sensor, and spraying a polyvinylpyrrolidone coating on the surface of the minimally invasive sensor by atomization.

[0016] In some embodiments, in one embodiment of the present invention, the minimally invasive sensor determines signals through the following steps:

[0017] Determining neural action potential signals and biochemical signals by collecting the frequency characteristics of the signals;

[0018] Determine the potentials corresponding to dopamine and acetylcholine by cyclic voltammetry, and determine the dopamine signal and the acetylcholine signal according to the potentials.

[0019] In some embodiments, in one embodiment of the present invention, the minimally invasive sensor measures disease signals through the following steps:

[0020] Analyze the spatio-temporal distribution characteristics of nerve action potential signals and biochemical signals, and extract signal characteristics related to diseases;

[0021] Based on the signal characteristics, construct a prediction model of signals and diseases through a machine learning model;

[0022] Perform disease prediction through the prediction model.

[0023] In some embodiments, in one embodiment of the present invention, the minimally invasive sensor measures disease signals through the following steps:

[0024] Analyze long-period nerve action potential signals and biochemical signals, and extract characteristic indicators related to diseases;

[0025] Perform whole transcriptome sequencing on the RNA expression of brain tissues at different stages of diseases, and perform correlation analysis on the significantly up-regulated or down-regulated mRNAs and the characteristic indicators to establish a spatio-temporal coupling model of signals and disease processes;

[0026] Perform disease prediction through the spatio-temporal coupling model.

[0027] In some embodiments, in one embodiment of the present invention, the coupling with zwitterionic polymers includes:

[0028] Use phosphocholine-based, carboxybetaine-based, sulfobetaine-based, polyphosphocholine-based, polysulfobetaine-based, and polycarboxybetaine-based zwitterionic polymers as exploration objects, and modify them on micro-needle electrodes respectively to characterize the wrapping of glial cells around the electrodes.

[0029] On the other hand, an embodiment of the present invention provides a signal acquisition system, including a minimally invasive sensor obtained by the implementation method of the minimally invasive sensor for neuroscience as described above, and a multi-channel signal acquisition circuit.

[0030] On the other hand, an embodiment of the present invention provides a disease prediction system, including the signal acquisition system as described above.

[0031] The embodiments of the present application at least include the following beneficial effects: The method provided by the embodiments of the present invention includes: preparing planar microneedles, preparing a nerve action potential sensing module on the front surface of the planar microneedles, and preparing a biochemical signal sensing module on the back surface of the planar microneedles; the biochemical signal sensing module includes a dopamine electrode and an acetylcholine electrode; stacking and integrating several pieces of the planar microneedles to obtain a minimally invasive sensor. The minimally invasive sensor prepared in the present application is a multifunctional microneedle array sensor, which can synchronously record nerve electrical signals and biochemical signals with high spatio-temporal resolution; and by stacking and integrating the planar microneedle arrays, the process difficulties of three-dimensional microneedle array microfabrication are alleviated, which is beneficial to reducing the preparation process of the minimally invasive sensor and improving the acquisition effect of the minimally invasive sensor. Description of the Drawings

[0032] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following introduces the drawings of the relevant technical solutions in the embodiments of the present invention or the prior art. It should be understood that the drawings introduced below are only for clearly expressing some embodiments of the technical solutions in the present invention for the convenience of those skilled in the art. Without creative efforts, other drawings can also be obtained based on these drawings.

[0033] Figure 1 It is a schematic flowchart of an embodiment of the implementation method of a minimally invasive sensor for neuroscience provided by the present invention;

[0034] Figure 2 It is a schematic structural diagram of an embodiment of a minimally invasive sensor for neuroscience provided by the present invention;

[0035] Figure 3 It is a schematic flowchart of another embodiment of the implementation method of a minimally invasive sensor for neuroscience provided by the present invention. Detailed Embodiments

[0036] The embodiments of the present invention are described in detail below. The examples of the embodiments are shown in the drawings, where the same or similar reference numerals represent the same or similar elements or elements with the same or similar functions throughout. The embodiments described below with reference to the drawings are exemplary and are only used to explain the present invention and should not be construed as a limitation of the present invention. For the step numbers in the following embodiments, they are only set for the convenience of explanation and no limitation is imposed on the order between the steps. The execution order of each step in the embodiments can be adaptively adjusted according to the understanding of those skilled in the art.

[0037] The field of biomedical engineering micro-devices (Bio-devices) is a highly interdisciplinary research field based on materials - micro-devices - biology. Since its development over the past 50-odd years, numerous remarkable achievements have been made in cutting-edge directions such as cell sensors, microfluidic devices, molecular detection chips, and wearable sensors. Bioinformatics sensing provides important technical means for revealing the mysteries of life and guiding disease treatment. Scientists have been eager to understand the nearly 100 billion neurons in the brain and find a series of new methods for treating neurological diseases such as Parkinson's syndrome, epilepsy, and Alzheimer's disease. The sensing and detection of physiological indicators of nerves, including neural electrical signals, biochemical signals, gene information, etc., is a key to unlocking the mystery of nerves and depends on gradually unfolding at different levels such as isolated cells, isolated tissues, and living animals. Neural microelectrode technology is a crucial tool for obtaining neural electrical signals.

[0038] However, there are still a series of problems that have not been overcome in the current sensing electrode technology for neuroscience. The microenvironment of cells and tissues is very complex; the size of cells (tens of micrometers) is in the microscopic range; and the cell membrane (6 nm thick) is an ultrathin and fragile biological barrier layer that is very easily damaged and affects cell activity. These microscopic characteristics make it particularly difficult to penetrate the cell membrane safely and efficiently. To meet the application requirements of the cell microenvironment, the miniaturization of key components of material devices is particularly important, but this also poses great challenges to the compatibility of complex functions such as sensing and treatment. Therefore, how to safely and efficiently detect and regulate the internal microenvironment of cells and tissues is a major problem in the development of contemporary biomedical engineering devices.

[0039] At the level of isolated cells, the recording of animal potentials of nerves is an important means of analyzing the information transmission of neural cell networks. Conventional multi-electrode array methods can only record extracellular electrical signals. Compared with recording electrical signals from the outside of the cell membrane, the detection of intracellular electrical signals based on the patch clamp technique can provide a more detailed cell signal peak spectrum, providing the possibility for accurately understanding neural cell activities. However, the patch clamp recording method has defects such as complex operation, difficulty in high-throughput recording of multiple cells, and affecting cell activity. The patch clamp is likely to affect cell activity and cannot be recorded for a long time. In previous studies, the applicant developed a method of assisting nanoneedles to penetrate the cell membrane based on electroporation and optoporation techniques, but the cell membrane usually heals within a few minutes and long-term recording is not possible. Whether there is a method for efficiently and long-term penetrating multiple neural cell membranes and the safety after long-term penetration of the cell membrane are all unknown scientific issues.

[0040] At the in vivo level, in addition to the action potential of nerve cells, biochemical signals of nerve cells, such as dopamine, acetylcholine, glutamate, etc., also closely affect and mark the progression of various neurological diseases. The sensing of these biochemical signals depends on an electrochemical three-electrode system and specific enzyme modification to achieve specific sensing. Due to the complex process of constructing and modifying precise electrochemical electrodes on the surface of three-dimensional microneedles, the sensing technology for high spatiotemporal resolution acquisition of biochemical signals in the brain has not been realized. Existing implantable brain electrodes mainly have the function of local electrical signal acquisition, but long-term acquisition of biochemical information in the brain has not been achieved. Due to the lack of a sensing electrode technology for high-resolution synchronous recording of neural electrical signals and biochemical signals, the spatiotemporal coupling relationship between neural electrical signals and biochemical signals in the brain has not been clarified.

[0041] The material selection, surface properties, geometric dimensions, and implantation operation of the invasive brain electrode are of great significance for the long-term implantation safety of the invasive electrode. Conventional rigid silicon-based or metal microelectrodes are prone to causing inflammatory reactions, tissue fibrosis, infection risks, etc. after implantation in the brain due to the differences in mechanical properties and material surface properties from brain tissue. These negative biocompatibility effects greatly affect the long-term stable recording of neural signals by current invasive electrodes. Especially for biochemical concentration signals that rely on signal amplitude recording, inflammatory reactions and tissue fibrosis are very likely to cause distortion of biochemical concentration signals.

[0042] In response to this, this application develops a new type of nanoneedle cell sensor device to solve the key difficulties in long-term recording of intracellular actions of nerve cells. The screening method is used to optimize the method of modifying cell-penetrating peptides, and a feasible method for the long-term penetration of nanoneedles into cells can be systematically found. Through gene sequencing, the safety of long-term penetration of cell membranes can be systematically revealed. Based on the modification of cell-penetrating peptides to extend the time effect of nanoneedles penetrating cell membranes, it can lay a necessary foundation for long-term and continuous detection of high-quality intracellular electrical signals.

[0043] This application creates a new type of multifunctional microneedle array sensor to solve the key difficulties in synchronously and highly spatiotemporally resolving the recording of neural electrical signals and biochemical signals. By using a mature microfabrication process to prepare fine micro-nano electrodes on a plane and then stacking and integrating the planar microneedle array into a three-dimensional microneedle array, the process difficulty of directly microfabricating the three-dimensional microneedle array is effectively solved. The high-precision and multifunctional synchronous sensing technology developed by the present invention has positive significance for revealing the scientific connotation of the spatiotemporal coupling between neural electrical signals and biochemical signals.

[0044] This application systematically elucidates the mechanical properties of implantable brain electrodes and the compatibility of material surface properties with long-term implantation in the brain, and can provide guiding principles for materials for long-term implantation of brain electrodes. Through two approaches, namely flexible electrode preparation and surface coating, the key problem of long-term implanted brain electrodes being wrapped by glial cells is overcome. By screening methods, the conditions for electrode flexibility and surface zwitterionic polymer modification are optimized, which is conducive to systematically finding solutions for long-term implantation of microneedle electrodes in the brain. The development of this technology has positive significance for revealing the spatio-temporal distribution law of neural electrical signals in the long term.

[0045] The implementation method and system of a minimally invasive sensor for neuroscience proposed according to an embodiment of the present invention will be described in detail below with reference to the accompanying drawings. First, the implementation method of the minimally invasive sensor for neuroscience proposed according to an embodiment of the present invention will be described with reference to the accompanying drawings.

[0046] Refer to Figure 1 , in an embodiment of the present invention, an implementation method of a minimally invasive sensor for neuroscience is provided, wherein the minimally invasive sensor is prepared through the following steps:

[0047] S100: Prepare planar microneedles, prepare a neural action potential sensing module on the front of the planar microneedles, and prepare a biochemical signal sensing module on the back of the planar microneedles; the biochemical signal sensing module includes a dopamine electrode and an acetylcholine electrode;

[0048] S200: Stack and integrate several pieces of the planar microneedles to obtain a minimally invasive sensor.

[0049] In some possible implementation manners, the neural action potential sensing module is used to collect neural action potential signals, the biochemical signal sensing module is used to collect biochemical signals, and the dopamine electrode and the acetylcholine electrode are respectively used to collect dopamine signals and acetylcholine signals. In this application, a minimally invasive sensor in the form of three-dimensional microneedles is prepared through stacking and integration.

[0050] Optionally, in an embodiment of the present invention, the preparing the planar microneedles, preparing a neural action potential sensing module on the front of the planar microneedles, and preparing a biochemical signal sensing module on the back of the planar microneedles includes:

[0051] Prepare planar microneedles by laser micro-cutting a polyimide substrate;

[0052] Prepare multi-channel platinum electrode micro-modules and micro-circuits on the front and back of the planar microneedles by photolithography;

[0053] Spin-coat a polyimide thin layer on the micro-circuit on the front to expose the front-end microelectrodes to obtain a neural action potential sensing module;

[0054] The microelectrodes on the back are covered by a mask, and acetylcholinesterase and Ag / AgCl are respectively modified to obtain a biochemical signal sensing module.

[0055] Optionally, in an embodiment of the present invention, the method further includes:

[0056] Perform oxygen plasma surface activation on the polyimide surface of the minimally invasive sensor and couple it with zwitterionic polymer to form a nanoscale surface coating.

[0057] In some possible implementation manners, since the zwitterionic polymer has hydrophilicity, it allows small molecules such as dopamine and acetylcholine to penetrate through the coating and contact the electrode surface for detection.

[0058] Optionally, in an embodiment of the present invention, the method further includes:

[0059] Optimize the coupling density of the surface coating of the minimally invasive sensor, and spray a polyvinylpyrrolidone coating on the surface of the minimally invasive sensor by atomization.

[0060] In some possible implementation manners, by spraying a polyvinylpyrrolidone coating on the surface, the surface hardness of the flexible microneedles is temporarily enhanced. After the microneedles penetrate the brain tissue, the surface PVP coating quickly dissolves in the brain tissue, exposing the electrode module of the flexible microneedles.

[0061] Optionally, in an embodiment of the present invention, the minimally invasive sensor determines the signal through the following steps:

[0062] Determine the nerve action potential signal and biochemical signal by collecting the frequency characteristics of the signal;

[0063] Determine the potentials corresponding to dopamine and acetylcholine by cyclic voltammetry, and determine the dopamine signal and acetylcholine signal according to the potentials.

[0064] In some possible implementation manners, this application uses the high-frequency characteristics of the action potential signal to distinguish from the biochemical signal; and uses cyclic voltammetry to explore the optimal potentials of dopamine and acetylcholine respectively to distinguish the dopamine signal from the acetylcholine signal; so as to realize the synchronous acquisition of electrical signals and biochemical signals.

[0065] Optionally, in an embodiment of the present invention, the minimally invasive sensor measures the disease signal through the following steps:

[0066] Analyze the spatio-temporal distribution characteristics of the nerve action potential signal and biochemical signal, and extract the signal characteristics related to the disease;

[0067] Based on the signal characteristics, construct a prediction model of the signal and the disease through a machine learning model;

[0068] Disease prediction is performed using the prediction model.

[0069] Optionally, in one embodiment of the present invention, the minimally invasive sensor measures the disease signal by the following steps:

[0070] Analyze long-period nerve action potential signals and biochemical signals to extract disease-related characteristic indicators;

[0071] Whole transcriptome sequencing of RNA expression in brain tissue at different stages of the disease is performed, and correlation analysis is performed between significantly upregulated or downregulated mRNA and the characteristic indicators to establish a spatiotemporal coupling model between the signal and the disease process;

[0072] Diseases are predicted using the spatiotemporal coupling model.

[0073] Optionally, in one embodiment of the present invention, coupling with a zwitterionic polymer comprises:

[0074] Phosphorylcholine, carboxylic acid betaine, sulfonic acid betaine, polyphosphorylcholine, polysulfonic acid betaine, and polycarboxylic acid betaine zwitterionic polymers were used as exploration objects and modified on microneedle electrodes respectively to characterize the wrapping of glial cells on the electrodes.

[0075] The implementation method provided by this application is described in detail below with a specific embodiment:

[0076] This application develops a technology for synchronous recording of potential and biochemical signals based on microneedle arrays: exploring innovative preparation methods for high-precision microneedle array sensors that can combine electrical and biochemical signal detection performance, achieving micro-involvement in the living brain, and synchronously detecting the spatiotemporal distribution of electrical and biochemical signals.

[0077] This application develops a stable sensing technology based on biocompatible flexible microneedle arrays: through two approaches, electrode flexibility and surface zwitterionic polymer coating, the positive effects of biocompatibility of microneedle brain electrodes are synergistically improved, micro-involvement in the brain is achieved, and electrical and biochemical signals can be stably recorded for a long time.

[0078] This application is based on the previous research work on micro-interventional sensing technology with multidisciplinary integration of "materials-microelectronics-biology", and further develops micro-interventional sensing technology for neuroscience. It is planned to tackle the two aspects of "microneedle array-based potential-biochemical signal synchronous recording technology" and "biocompatible flexible microneedle array stable sensing technology", focusing on solving the difficult problems of "long-term", "diversified" and "more stable" acquisition of neural signals. Then, Parkinson's syndrome is used as a representative neurological disease object to explore biological mechanisms and evaluate drugs. Obtain high-content physiological electrical signals and biochemical signals related to Parkinson's disease at the in vivo level, revealing the spatiotemporal coupling mechanism of signals and disease progression, as well as the drug feedback mechanism.

[0079] Specifically, the present application develops a potential-biochemical signal synchronous recording technology based on a microneedle array:

[0080] Referring to Figure 2 As shown, the present invention prepares a nerve action potential and biochemical signal (dopamine, acetylcholine) sensing module on the front and back of the planar microneedles respectively, so as to solve the mutual interference between the electrical signal electrode and the biochemical signal electrode during the preparation process; and then integrates 5 planar microneedles into a three-dimensional microneedle array to solve the problem that it is difficult to directly process three-dimensional microneedles.

[0081] Referring to Figure 3 As shown, first, a planar microneedle (including 5 microneedles, each with a diameter of 300 microns and a length of 800 microns) array is prepared by laser micro-cutting a polyimide substrate. High-precision (10-micron precision) multi-channel platinum electrode micro-modules and micro-circuits are respectively prepared on the front and back of the planar microneedles by photolithography. A polyimide thin layer is spin-coated on the microelectrode circuit on the front for insulation, and only the front-end microelectrode is exposed as a sensing module for recording the action potential of nerve cells. The microelectrodes on the back are covered with a mask and are respectively modified with acetylcholinesterase (for specifically detecting acetylcholine) and Ag / AgCl (as a reference electrode) as a sensing module for detecting biochemical signals. Then, 5 planar microneedle arrays are stacked and integrated into a 5×5 three-dimensional array. Since the microneedles are distributed in a three-dimensional array, it has the advantage of recording the spatial distribution of nerve signals.

[0082] The present application develops a stable sensing technology based on a biocompatible flexible microneedle array:

[0083] Based on the high-precision microneedle array sensor developed in the previous work and having both electrical signal and biochemical signal detection performance, the present invention mainly improves the biocompatibility of the electrode from two ways: flexible preparation of the electrode and surface coating, so as to achieve long-term implantation in the brain for signal recording. Further explore how to optimize the mechanical hardness of the microneedles to reduce the biocompatibility problem caused by the difference in mechanical hardness between the microneedle electrodes and the brain tissue after the microneedle electrodes penetrate into the brain tissue; and then explore how to construct a biocompatible coating on the surface of the microneedle electrodes to inhibit the non-specific adsorption of proteins in the brain tissue on the electrode surface, so as to achieve the effect of reducing the inflammatory response and tissue fibrosis. Through the improvement of both the mechanical properties and surface properties of the microneedles, it is expected to realize more stable implantation of the electrodes in the brain for stable recording of physiological signals.

[0084] 1) A flexible micro-needle array is fabricated using a flexible polyimide film substrate, and substrates with different thicknesses (2 - 20 microns thick) are selected to control the flexibility of the electrodes. The polyimide surface of the micro-needle array sensor is activated by oxygen plasma and coupled with zwitterionic polymers to form a nanoscale surface coating. Since zwitterionic polymers are hydrophilic, small molecules such as dopamine and acetylcholine can pass through the coating and contact the electrode surface for detection.

[0085] 2) Electrodes with different degrees of flexibility are explored and implanted in the animal brain for one month, and then the electrodes are taken out to characterize the wrapping of glial cells around the electrodes; in terms of surface modification of zwitterionic polymers, phosphocholine-based, carboxybetaine-based, sulfobetaine-based, polyphosphocholine-based, polysulfobetaine-based, and polycarboxybetaine-based zwitterionic polymers are used as exploration objects and are respectively modified on the micro-needle electrodes. After being implanted in the animal brain for one month, the electrodes are taken out to characterize the wrapping of glial cells around the electrodes. Through these studies, the optimal conditions for the flexibility of the electrodes and surface modification are explored.

[0086] 3) The coupling density of the surface coating is further optimized to maintain the sensing performance of the micro-needle electrodes in detecting electrical signals and biochemical signals. Furthermore, a water-soluble polyvinylpyrrolidone (PVP) coating is sprayed on the micro-needle surface by atomization to temporarily enhance the surface hardness of the flexible micro-needles. After the micro-needles penetrate the brain tissue, the surface PVP coating quickly dissolves in the brain tissue, exposing the electrode module of the flexible micro-needles.

[0087] Example 1:

[0088] Micro-interventional sensing research for neuroscience, instrument system for in vivo animal experiment applications

[0089] Explore the methods and conditions for modifying various cell-penetrating peptides on the surface of nano-needle electrodes:

[0090] After evaluating 18 different cell-penetrating peptides modified on nano-needle electrodes, the transmembrane efficiency, transmembrane duration, and transmembrane stability of the nano-needles to cell membranes are evaluated. The spacing of the nano-needles is optimized so that the bottom of the nerve cell body can be effectively covered by multiple nano-needles, and the optimal state of the nano-needles penetrating the cell membrane is further determined. Using whole-transcriptome mRNA sequencing, it is clarified whether the cell-penetrating peptide-modified nano-needle device affects cell mRNA expression. It is clarified whether the cell-penetrating peptide-modified nano-needles affect cell activities, proliferation, metabolism, electrical signals, pacing, etc. after long-term penetration of the cell membrane. After evaluating the nano-needles penetrating the cell membrane, the sensitivity, signal-to-noise ratio, ion channel signal peak resolution, persistence, and accuracy of the nano-needle sensor in long-term recording of intracellular electrical signals are evaluated.

[0091] Simultaneous recording of potential-biochemical signals based on micro-needle arrays:

[0092] Using Parkinson's disease as a model, the prepared microneedle sensors were inserted into the brains of live rats. Evaluate the stability of the microneedle array fixed in the brain and the safety of brain tissues. Distinguish using the high-frequency characteristics of action potential signals and biochemical signals; and use cyclic voltammetry to explore the optimal potentials of dopamine and acetylcholine respectively to distinguish dopamine signals from acetylcholine signals. Evaluate the sensitivity, signal-to-noise ratio, persistence, and accuracy of the microneedle sensors in synchronously recording action potentials and biochemical signals in vivo.

[0093] Reveal the spatio-temporal coupling relationship between the neuroelectrical signals of Parkinson's disease and dopamine and acetylcholine signals, and test the effects of three representative Parkinson's drugs, such as levodopa, carbidopa, and selegiline, on neurophysiological signals. Analyze the spatio-temporal distribution characteristics of electrical signals and biochemical signals, extract signal feature indicators associated with the disease model, and select appropriate machine learning models for construction, such as support vector machine (SVM), random forest (RF), or neural network (NN), and select appropriate models according to the data type and distribution characteristics of the features. To ensure the generalization ability and stability of the model, we will adopt the method of cross-validation, divide the data set into training set, validation set, and test set, and optimize and adjust the model parameters. During the training process, multiple indicators are selected to comprehensively evaluate the model performance, and visual tools such as ROC curves and confusion matrices are used to intuitively present the model performance. In terms of feature selection, methods such as correlation analysis and principal component analysis (PCA) are used to screen out the feature combinations with the strongest correlation with the disease model. In addition, sensitivity analysis will also be used to evaluate the impact of each feature on the model prediction results and explore its potential biological significance. Finally, a visualization interface and decision rules of the model will be constructed and applied to the diagnosis, staging, and personalized formulation of treatment plans for Parkinson's disease patients to establish a disease evaluation model.

[0094] Stable Sensing Based on Biocompatible Flexible Microneedle Arrays:

[0095] Using Parkinson's disease as a model, the prepared flexible microneedle sensors were inserted into the brains of live rats. Explore the long-term (more than one month) stability of the flexible microneedles fixed in the brain and the safety of brain tissues. Evaluate the improvement effect of the sensitivity, stability, and accuracy of the flexible microneedle sensors in long-term recording of action potentials and biochemical signals in vivo. Furthermore, use the microneedle sensors to evaluate their performance in long-term (continuously for more than one month) synchronous recording of action potentials and biochemical signals in the live brain, as well as the spatio-temporal distribution of long-term recordings.

[0096] Analyze long-period electrical signals and biochemical signals, and extract signal feature indicators corresponding to disease models. Perform whole-transcriptome sequencing on the RNA expression of brain tissues at different stages of Parkinson's disease, and conduct correlation analysis on the significantly up-regulated or down-regulated mRNAs with the significant features of electrical signals and biochemical signals to reveal the spatio-temporal coupling mechanism between signals and the disease process.

[0097] Analyze long-period neural electrical signals and biochemical signals, and extract their significant features, such as changes at different time points, different frequency components, and different waveforms. For the RNA sequencing results, identify significantly differentially expressed genes (DEGs) at different stages of Parkinson's disease (such as early, middle, and late stages), and focus on up-regulated or down-regulated mRNAs. Then, use a variety of statistical methods for correlation analysis, including correlation analysis, regression analysis, clustering analysis, network analysis, and time series analysis. Reveal potential causal relationships and construct a spatio-temporal coupling model for the pathogenesis of Parkinson's disease. Comprehensively use a variety of analysis strategies to deeply understand the complex dynamic relationships among neural electrical signals, biochemical signals, and gene expression in Parkinson's disease, and clarify the roles and mechanisms of signal changes and the disease process in the spatio-temporal dimension.

[0098] Further combine multi-channel weak signal amplification and detection technology, develop a multi-channel high-frequency and low-frequency signal acquisition circuit based on FPGA, and instrument this into a micro-needle electrode array to form a set of instrument systems for in vivo animal experiment applications.

[0099] In summary, the method provided in the embodiments of the present application includes: preparing planar micro-needles, preparing a neural action potential sensing module on the front of the planar micro-needles, and preparing a biochemical signal sensing module on the back of the planar micro-needles; the biochemical signal sensing module includes a dopamine electrode and an acetylcholine electrode; stacking and integrating several pieces of the planar micro-needles to obtain a minimally invasive sensor. The minimally invasive sensor prepared in the present application is a multi-functional micro-needle array sensor that can synchronously record neural electrical signals and biochemical signals with high spatio-temporal resolution; and by stacking and integrating the planar micro-needle array, it alleviates the process difficulties of three-dimensional micro-needle array microfabrication, which is beneficial to reducing the preparation process of the minimally invasive sensor and improving the acquisition effect of the minimally invasive sensor.

[0100] On the other hand, an embodiment of the present invention provides a signal acquisition system, including a minimally invasive sensor obtained by the implementation method of the minimally invasive sensor for neuroscience as described above, and a multi-channel signal acquisition circuit.

[0101] On the other hand, an embodiment of the present invention provides a disease prediction system, including the signal acquisition system as described above.

[0102] Similarly, the content in the above method embodiments is applicable to the system embodiments. The functions specifically implemented by the system embodiments are the same as those in the above method embodiments, and the beneficial effects achieved are also the same as those in the above method embodiments.

[0103] In some alternative embodiments, the functions / operations mentioned in the block diagrams may not occur in the order mentioned in the operation diagrams. For example, depending on the functions / operations involved, two consecutive blocks shown may actually be executed substantially simultaneously or the blocks can sometimes be executed in the reverse order. Additionally, the embodiments presented and described in the flowcharts of the present invention are provided by way of example for the purpose of providing a more comprehensive understanding of the technology. The disclosed methods are not limited to the operations and logical flows presented herein. Alternative embodiments are contemplated where the order of various operations is changed and where sub-operations described as part of a larger operation are executed independently.

[0104] Furthermore, although the present invention has been described in the context of functional modules, it should be understood that, unless otherwise stated to the contrary, one or more of the functions and / or features may be integrated in a single physical device and / or software module, or one or more functions and / or features may be implemented in separate physical devices or software modules. It can also be understood that a detailed discussion of the actual implementation of each module is not necessary for understanding the present invention. Rather, given the attributes, functions, and internal relationships of the various functional modules in the devices disclosed herein, the actual implementation of the modules will be understood within the ordinary skill of an engineer. Thus, those skilled in the art can implement the present invention as set forth in the claims without undue experimentation. It can also be understood that the specific concepts disclosed are merely illustrative and are not intended to limit the scope of the present invention, which is determined by the full scope of the appended claims and their equivalents.

[0105] The logic and / or steps represented in the flowcharts or otherwise described herein, for example, can be considered as a sequenced list of executable instructions for implementing logical functions, and can be specifically embodied in any computer-readable medium for use by or in connection with a program execution system, apparatus, or device, such as a computer-based system, a system including a processor, or other systems that can fetch and execute the program from the program execution system, apparatus, or device. For the purposes of this specification, a "computer-readable medium" can be any device that can contain, store, communicate, propagate, or transport the program for use by or in connection with the program execution system, apparatus, or device.

[0106] In the foregoing description of this specification, the descriptions referring to the terms "one embodiment / example", "another embodiment / example" or "certain embodiments / examples", etc. mean that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0107] Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the claims and their equivalents.

[0108] The above is a specific description of the preferred embodiments of the present invention, but the present invention is not limited to the described embodiments. Those skilled in the art can also make various equivalent deformations or substitutions without departing from the spirit of the present invention, and these equivalent deformations or substitutions are all included in the scope defined by the claims of the present invention.

Claims

1. A method for implementing a minimally invasive sensor for neuroscience, characterized in that: The minimally invasive sensor was prepared by the following steps: Prepare a planar microneedle, prepare a neural action potential sensing module on the front side of the planar microneedle, and prepare a biochemical signal sensing module on the back side of the planar microneedle; The biochemical signal sensing module includes a dopamine electrode and an acetylcholine electrode; Several sheets of the planar microneedles are stacked and integrated to obtain a minimally invasive sensor.

2. The method for implementing a minimally invasive sensor for neuroscience according to claim 1, characterized in that: The method of preparing a planar microneedle, preparing a neural action potential sensing module on the front side of the planar microneedle, and preparing a biochemical signal sensing module on the back side of the planar microneedle comprises: Planar microneedles were prepared by laser microcutting of polyimide substrates; By photolithography, multi-channel platinum electrode micromodules and microcircuits are prepared on the front and back sides of the planar microneedles; A polyimide thin layer is suspended on the front microcircuit to expose the front microelectrode to obtain a neural action potential sensing module; The microelectrodes on the back are covered with a mask to modify acetylcholinesterase and Ag / AgCl respectively to obtain a biochemical signal sensing module.

3. The method for implementing a minimally invasive sensor for neuroscience according to claim 2, characterized in that: The method further comprises: The polyimide surface of the minimally invasive sensor is activated by oxygen plasma and coupled with a zwitterionic polymer to form a nanoscale surface coating.

4. The method for implementing a minimally invasive sensor for neuroscience according to claim 1, characterized in that: The method further comprises: The coupling density of the coating on the surface of the minimally invasive sensor is optimized, and a polyvinyl pyrrolidone coating is sprayed on the surface of the minimally invasive sensor by atomization.

5. The method for implementing a minimally invasive sensor for neuroscience according to claim 1, characterized in that: The minimally invasive sensor determines the signal by the following steps: By collecting the frequency characteristics of the signal, the nerve action potential signal and biochemical signal are determined; The potentials corresponding to dopamine and acetylcholine are determined by cyclic voltammetry, and the dopamine signal and the acetylcholine signal are determined based on the potentials.

6. The method for implementing a minimally invasive sensor for neuroscience according to claim 1, characterized in that: The minimally invasive sensor measures disease signals through the following steps: Analyze the spatiotemporal distribution characteristics of nerve action potential signals and biochemical signals to extract disease-related signal features; Based on the signal characteristics, a prediction model of signals and diseases is constructed through a machine learning model; Disease prediction is performed using the prediction model.

7. The method for implementing a minimally invasive sensor for neuroscience according to claim 1, characterized in that: The minimally invasive sensor measures disease signals through the following steps: Analyze long-period nerve action potential signals and biochemical signals to extract disease-related characteristic indicators; Whole transcriptome sequencing of RNA expression in brain tissue at different stages of the disease is performed, and correlation analysis is performed between significantly upregulated or downregulated mRNA and the characteristic indicators to establish a spatiotemporal coupling model between the signal and the disease process; Diseases are predicted using the spatiotemporal coupling model.

8. The method for implementing a minimally invasive sensor for neuroscience according to claim 3, characterized in that: The coupling with the zwitterionic polymer comprises: Phosphorylcholine, carboxylic acid betaine, sulfonic acid betaine, polyphosphorylcholine, polysulfonic acid betaine, and polycarboxylic acid betaine zwitterionic polymers were used as exploration objects and modified on microneedle electrodes respectively to characterize the wrapping of glial cells on the electrodes.

9. A signal acquisition system, characterized in that: The invention comprises a minimally invasive sensor obtained by the method for realizing a minimally invasive sensor for neuroscience as described in any one of claims 1 to 8, and a multi-channel signal acquisition circuit.

10. A disease prediction system, characterized in that: Comprising the signal acquisition system as claimed in claim 9.

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