An electrically coupled chip system for single cell and substrate interaction studies

Abstract

The disclosure provides a power coupling chip system for single cell and substrate interaction research, and relates to the technical field of cell detection. The system comprises a first subsystem and a second subsystem, each of which comprises a culture module, an observation module and an electrical signal transmission module, and the system further comprises a post-processing module; the hydrogel is used for carrying and culturing cells; the electrical signal transmission module comprises a PCB circuit board and a transmission electrode for transmitting an electrical stimulation signal; the observation module comprises a shooting device which collects images of the culture chamber during the electrical stimulation process through a transparent glass; the post-processing module is used for post-processing the images obtained by each subsystem to obtain the morphology and behavior information of the cells. The disclosure can more comprehensively obtain the kinetic information of the cells under the action of the coupled physical signals by actively stimulating and passively stimulating the cells through the stiffness-adjustable and electrical-adjustable hydrogel.

Description

Technical Field

[0001] This disclosure relates to the field of cell detection technology, and in particular to an electrically coupled chip system for studying single-cell-substrate interactions. Background Technology

[0002] As the basic unit of life, the cell's complex dynamic behavior is closely linked to the normal functioning of the organism. Cellular dynamics, especially cell migration, are key processes in the development of various physiological and pathological conditions, such as wound healing, embryonic development, inflammatory responses, and tumor metastasis. Cells do not exist in isolation within the body's microenvironment but constantly sense and respond to various signals within it.

[0003] In recent years, the role of mechanical signals in regulating cellular behavior has received increasing attention. Cells can not only generate traction forces to actively change their shape and migrate, but also be passively influenced by mechanical forces through interactions with the extracellular matrix. Similarly, electrical signals can actively respond to exogenous electrical signals and interact with the matrix microenvironment through physiological electrical signals, thereby providing feedback on cellular life activities. Therefore, understanding the active and passive response behaviors of single cells and the substrate is crucial for comprehensively revealing the mechatronic mechanisms of cells in real physiological environments. However, systematic research on how electrically coupled signals in the cellular microenvironment regulate cellular dynamics is still lacking. Summary of the Invention

[0004] In view of this, the purpose of this disclosure is to propose an electrically coupled chip system for studying single-cell-substrate interactions, which can specifically address existing problems.

[0005] Based on the above objectives, in a first aspect, this disclosure proposes an electrically coupled chip system for studying single-cell-substrate interactions, comprising: the system including a first subsystem and a second subsystem, each subsystem including a culture module, an observation module, and an electrical signal transmission module; the system further including a post-processing module; the culture module including a cell culture chamber comprising a transparent glass and a hydrogel placed on the transparent glass, the hydrogel being used to support and culture cells; the hydrogel of the first subsystem being a conductive hydrogel, and the hydrogel of the second subsystem being a non-conductive hydrogel; the conductive hydrogel being used to receive electrical coupling to induce a passive response in the cells, and the cells supported by the non-conductive hydrogel being used to directly receive electrical coupling to induce an active response in the cells; the electrical signal transmission module including a PCB circuit board for transmitting electrical stimulation signals and transmission electrodes; the observation module including an imaging device that acquires images of the culture chamber during the electrical stimulation process through the transparent glass; and the post-processing module for post-processing the images obtained from each subsystem to obtain morphological and behavioral information of the cells.

[0006] Secondly, a method for generating cell information is also provided, which employs the power coupling chip system described in the first aspect. The method includes generating kinetic behavior information of cells through the power coupling chip system.

[0007] Thirdly, an electronic device is also provided, including a memory, a processor, and a computer program stored in the memory and executable on the processor, the processor running the computer program to implement the method of the second aspect.

[0008] Fourthly, a computer-readable storage medium is also provided, on which a computer program is stored, the computer program being executed by a processor to implement the method described in the second aspect.

[0009] Fifthly, a computer program product is also provided, comprising a computer program that is executed by a processor to implement the method described in the second aspect.

[0010] In summary, this disclosure has at least the following beneficial effects: by coupling mechanical and electrical signals in cells, multidimensional coupled signal analysis of the active and passive responses of cells can be performed, thereby obtaining more comprehensive dynamic information of cells under the action of multiple physical fields, which helps to better understand the mechanobiological mechanisms of cells. Attached Figure Description

[0011] In the accompanying drawings, unless otherwise specified, the same reference numerals throughout the various drawings denote the same or similar parts or elements. These drawings are not necessarily drawn to scale. It should be understood that these drawings depict only some embodiments disclosed in this disclosure and should not be construed as limiting the scope of this disclosure.

[0012] Figure 1 A schematic diagram of a power-coupled chip system structure for studying single-cell-substrate interactions according to an embodiment of the present disclosure is shown. Figure 2 A schematic diagram illustrating the principle of a power-coupled chip system according to an embodiment of the present disclosure for quantifying single-cell-substrate interactions is shown. Figure 3 The fabrication process and characterization results of the cell-electrically coupled chip system according to embodiments of this disclosure are shown; Figure 4 The design and electromechanical characterization results of cellular conductive / non-conductive hydrogel substrates according to embodiments of this disclosure are shown. Figure 5 The results of cell dynamics information inversion based on an electrically tunable and stiffness-tunable hydrogel substrate according to an embodiment of the present disclosure are shown. Figure 6 The deformation characterization results of an ion-conductive hydrogel substrate according to an embodiment of the present disclosure are shown. Detailed Implementation

[0013] The present disclosure will now be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention. Furthermore, it should be noted that, for ease of description, only the parts relevant to the invention are shown in the accompanying drawings.

[0014] It should be noted that, unless otherwise specified, the embodiments and features described in this disclosure can be combined with each other. This disclosure will now be described in detail with reference to the accompanying drawings and embodiments.

[0015] Figure 1This disclosure illustrates an electrically coupled chip system for studying single-cell-substrate interactions. In embodiments of this disclosure, the system includes a first subsystem and a second subsystem. Each subsystem includes a culture module, an observation module, and an electrical signal transmission module. The system also includes a post-processing module. The culture module includes a cell culture chamber comprising a transparent glass and a hydrogel placed on the transparent glass. The hydrogel is used to support and culture cells. The hydrogel in the first subsystem is conductive, while the hydrogel in the second subsystem is non-conductive. The conductive hydrogel receives electrically coupled signals to induce a passive response in the cells, while the cells supported by the non-conductive hydrogel directly receive electrically coupled signals to induce an active response. The electrical signal transmission module includes a PCB circuit board for transmitting electrical stimulation signals and transmission electrodes. The observation module includes an imaging device that acquires images of the culture chamber during the electrical stimulation process through the transparent glass. The post-processing module is used to post-process the images obtained from each subsystem to obtain cell dynamics information.

[0016] The imaging can be performed on the upper surface of the hydrogel. The captured images include corresponding images from both the first and second subsystems. The first subsystem produces images of cells receiving a passive response, while the second subsystem produces images of cells receiving an active response.

[0017] This embodiment discloses the interaction between a single cell and a substrate based on a power-coupled chip system.

[0018] This disclosure utilizes electrically tunable and stiffness-tunable hydrogel substrates to perform active and passive response analysis on cells, enabling a more comprehensive acquisition of cellular dynamic information, including dynamic behavior information, under the coupling of multiple physical fields, which helps to better understand the mechanobiological mechanisms of cells.

[0019] In some optional implementations of any embodiment of this disclosure, the image includes an image before deformation and an image after deformation; the imaging device is a high-speed fluorescence microscope; and the dynamic behavior information includes a displacement field and a traction field. The high-speed fluorescence microscope is used to capture images of single cells in the culture chamber in real time before the electrocoupling action, and to obtain an image before deformation. After the electrocoupling action, the microscope captures images of single cells in the culture chamber in real time, and to obtain an image after deformation. The post-processing module is further used to perform inversion calculations on the captured images to obtain the displacement field and traction field of the cell in response to the electrocoupling signal.

[0020] The image before deformation includes at least one image, and the image after deformation includes at least one image.

[0021] These methods utilize high-speed fluorescence microscopy to capture instantaneous dynamic changes in cells during electrical stimulation, avoiding image blurring. By analyzing the changes in images before and after deformation, visual images are transformed into precise data on physical changes, which helps to obtain more accurate information on dynamic behavior.

[0022] Optionally, fluorescent particles are present on the surface of the hydrogel. Under the action of an electrical coupling signal, the cell's response is transmitted to the hydrogel substrate and manifested through the displacement difference of the fluorescent particles. The post-processing module is further used to analyze the position of fluorescent particles in the images acquired by each subsystem for inversion calculation.

[0023] These alternative implementations can precisely calculate the minute forces exerted by cells on the substrate by tracking minute changes in particle position, significantly improving the sensitivity and accuracy of mechanobiological detection.

[0024] Optionally, the gold electrode and the chromium electrode can be square or round; the PCB circuit board is a square printed circuit board, and the PCB pads of the PCB circuit board are provided with multiple electrode leads. The hollow shape and thickness of the inner ring of the PCB circuit board are consistent with the outer contour shape and thickness of the gold electrode; the electrodes of the gold electrode located around the transparent glass are connected to the PCB pads through conductive silver paste, and the adapter is used to receive external electrical signals.

[0025] The conductive silver paste bonding process in these optional implementations is simple and has low resistance, ensuring efficient and stable transmission of electrical signals from the PCB board to the glass chip. The inner ring of the PCB board conforms to the contour of the gold electrodes. This design enables seamless docking between the microfluidic chip and the macroscopic circuit board, ensuring both a wide field of view for microscopic observation and a reliable electrical interface.

[0026] In some optional implementations of any embodiment of this disclosure, the PCB circuit board includes PCB pads, adapters, and PCB electrodes; the transmission electrodes include chromium electrodes and gold electrodes, the chromium electrodes are disposed on the upper surface of the transparent glass, the gold electrodes are disposed above the chromium electrodes, and a closed loop between the electrodes is formed in the incubation chamber, the gold electrodes, and the PCB electrodes; the gold electrodes include symmetrical electrodes and asymmetrical electrodes, the symmetrical electrodes are used to apply electrical stimulation with a uniform electric field, and the asymmetrical electrodes are used to apply electrical stimulation with a non-uniform electric field.

[0027] In these implementations, the chromium layer acts as a "primer," increasing the bonding strength between the gold electrode and the glass, while the gold layer ensures excellent conductivity. The fusion design of symmetrical and asymmetrical electrodes allows the system to not only apply electrical stimulation but also study the differential responses of cells under different electric field modes, both uniform and non-uniform.

[0028] In some optional implementations of any embodiment of this disclosure, the system further includes an electrode generation module; the electrode generation module is used to generate chromium electrodes and gold electrodes in the transport electrode using photolithography and magnetron sputtering techniques according to the set electrode size, electrode shape, array size and array density.

[0029] These implementation methods enable flexible electrode fabrication according to different application requirements, resulting in electrodes with different array densities or shapes, thus improving the versatility of the chip and the repeatability of experiments.

[0030] In some optional implementations of any embodiment of this disclosure, the ion concentration, stiffness, and voltage of the electrical stimulation signal of the hydrogel are preset variables, which are related to the magnitude of the cell response during the electrocoupling process of the hydrogel.

[0031] These implementations set multiple parameters as variables and can also be combined with tunable material systems, enabling the study of a single parameter or the combined effects of multiple factors coupled together, for the entire system.

[0032] In some optional implementations of any embodiment of this disclosure, the conductive hydrogel is an acrylamide and sodium methacrylamide / PEDOT:PSS gel, wherein the combination of acrylamide and PEDOT:PSS constitutes an electronically conductive hydrogel, the combination of acrylamide and sodium methacrylamide constitutes an ionically conductive hydrogel, and the non-conductive hydrogel is a polyacrylamide gel.

[0033] The PEDOT:PSS combination in these implementations provides electronic conductivity, facilitating the accurate detection of passive responses on electronically conductive substrates. The AlgMA combination provides ionic conductivity, enabling the accurate study of electric field-induced hydrogel deformation. Polyacrylamide, as a purely mechanical substrate, helps in the accurate detection of active cell responses to direct electrical stimulation. Furthermore, the use of these biocompatible materials ensures cell viability during experiments.

[0034] This disclosure aims to precisely quantify and analyze the active and passive response behaviors of cells mediated by electrocoupling signals and elucidate their role in mechanobiology. First, a stable cell electrocoupling chip system is constructed using photolithography and magnetron sputtering techniques. Electrical and mechanical signals are loaded by fabricating safe and reliable gold electrode patterns on transparent glass. Using non-conductive polyacrylamide hydrogel (PAAm), the active response behavior of cells under electrocoupling is studied. Further, PEDOT:PSS is introduced to form an electronically conductive hydrogel, and sodium methacryloyl alginate (AlgMA) is introduced to form an ionically conductive hydrogel to simulate the passive response behavior of cells under electrocoupling. By introducing conductive / non-conductive cellular microenvironments, the direct and indirect effects of electrical and mechanical signals on single cells are quantified, respectively. Finally, traction force microscopy (TFM) is used to analyze the acquired images, and the cell displacement changes and mechanical spectra mediated by electrocoupling signals are obtained through inversion and quantification. By combining the analysis of active and passive cell responses, this disclosure will provide new insights into the mechanobiological mechanisms of cells under the influence of electrically coupled signals, and provide technical references for the development of cell therapies and tissue engineering based on the regulation of multi-physical signal coupling, such as accelerating wound healing and promoting functional recovery in nerve repair.

[0035] This disclosed power-coupled chip system is a cell microenvironment system based on glass-gold electrodes, designed to investigate the kinetics of cell responses under electrocoupling. The highly transparent glass enables real-time cell observation, while the gold electrodes ensure stable transmission of cellular electrical signals. The gold electrodes consist of two pairs: a pair of symmetrical electrodes and a pair of asymmetrical electrodes, allowing simultaneous study of cell dynamics under both uniform and non-uniform electric fields. In cell culture using this power-coupled chip system, a custom-designed glass ring is assembled to a glass substrate using biosafety polydimethylsiloxane (PDMS). The electrode assembly includes a working electrode and lead electrodes; one end of the working electrode is circular or square. The lead electrodes are connected to electrode soldering areas on a PCB circuit board via soldering areas on the outer side of the glass. A closed loop is formed between the electrodes in the culture chamber, the glass-gold electrodes, and the PCB electrodes. The electrode soldering areas on the PCB circuit board are exposed, enabling rapid assembly with the glass-gold electrodes. The lead electrodes on the PCB are encapsulated with polyimide (PI) material to prevent damage or even open circuits during experiments. The entire system, under the influence of an external electrical signal and substrates of varying stiffness, transmits electrical and mechanical signals to the cell culture chamber to physically stimulate cells. Simultaneously, confocal microscopy is used to capture and image the dynamic cell response during electrical stimulation in real time. Finally, TFM is used to calculate the displacement and traction fields of single cells under electrical coupling. The active and passive responses of single cells are achieved using hydrogels with different substrate properties. Non-conductive hydrogels (polyacrylamide gels) are used to characterize the active cell response to electrical stimulation, while conductive hydrogels (acrylamide and PEDOT:PSS gels, or acrylamide and methacryloylated sodium alginate gels) are used to characterize the passive cell response induced by the hydrogel's response to electrical stimulation. Fluorescent particles are added to the prepolymer solution before gelation for both types of gels. During gelation, the fluorescent particles are collected on the gel surface using an inverted method, and the gel surface is modified with proteins. Subsequently, as the cells stretch the substrate, the position of the fluorescent particles changes accordingly. Further inversion calculations can be performed by acquiring images of the fluorescent particles before and after processing. Based on this method, the dynamic behavior of single cells mediated by multiple physical coupling signals can be characterized. It not only collects the dynamic response mediated by cell electrical coupling from a macroscopic perspective, but also characterizes the physical signals, namely mechanical signals and displacement information, in the cell dynamic response process from a microscopic perspective.

[0036] The construction of the second subsystem, namely the cell-active electrical coupling response system: Cellular active electrocoupling response is achieved through a stiffness-tunable and non-conductive substrate. The preparation process of polyacrylamide gels with different stiffnesses includes the following steps: A 40% (w / v) aqueous solution of acrylamide (AM) monomer and a 2% (w / v) aqueous solution of bis-acrylamide (Bis) were prepared using deionized water as stock solutions. Adjusting the ratio of the two stock solutions yielded polyacrylamide gels with different stiffnesses. First, 1 μL / mL of red fluorescent particles was added to the prepolymer solution, and the mixture was vortexed for 2 minutes. Then, 5 μL / mL of 10% ammonium persulphate (APS) and 10 μL / mL of 5% tetramethylethylenediamine (TEMED) solution were added. The mixture was rapidly vortexed for approximately 10 seconds, and 30 μL of the hydrogel liquid was quickly dropped onto the surface of a modified culture dish. A prepared coverslip was then quickly placed over the hydrogel surface, and the dish was immediately inverted to allow the fluorescent particles to deposit onto the hydrogel surface. After 30 minutes of polymerization, add 1 mL of deionized water to the culture dish, gently remove the coverslip with tweezers, and check the integrity of the gel substrate. Finally, add deionized water to the culture dish containing the polyacrylamide gel substrate and set aside for later use.

[0037] The construction of the first subsystem, namely the cellular passive electrical coupling response system: Electron-conductive hydrogels achieve effective stimulation intensity at a cell-safe voltage under the influence of an applied voltage. Specifically, for hydrogels with high conductivity, current can more easily penetrate the hydrogel, enabling cells to obtain an effective passive response.

[0038] The substrate preparation process for electronically conductive hydrogels includes the following steps: A electrically tunable cell matrix was prepared by incorporating PEDOT:PSS into a polyacrylamide (PAAm) hydrogel. The PAAm and PEDOT:PSS in this hydrogel formed an interpenetrating polymer network with good biocompatibility and electronic conductivity. First, the PEDOT:PSS stock solution was pretreated with 5% (v / v) dimethyl sulfoxide (DMSO) and sonicated for 10 minutes. DMSO treatment reshaped the microstructure of PEDOT:PSS, promoting the separation of the insulating PSS phase and enabling the conductive PEDOT phase to form a denser, interconnected nanofiber network. Subsequently, different volumes of pretreated PEDOT:PSS (final concentrations of 0.1% or 1%) were added to a pre-prepared polyacrylamide gel precursor solution to obtain hydrogels with different conductivity. Furthermore, the addition of 0.1% Triton X-100 to the PAAm-PEDOT:PSS mixed solution effectively prevented the aggregation of PEDOT:PSS nanofibers during polymerization, ensuring a more uniform distribution within the gel network. After gelation, this cross-linked conductive hydrogel requires post-treatment: first, it is soaked in 5% (volume ratio) DMSO for 10 hours to further optimize the internal conductive network; then, it is soaked in deionized water for 24 hours, with the water changed every 6 hours, to remove unreacted monomers, initiator fragments and soluble oligomers.

[0039] The substrate preparation process for ion-conducting hydrogels includes the following steps: Based on the conductivity and deformability of hydrogels, an interpenetrating network structure formed by polyacrylamide (PAAm) and methacrylated alginate (AlgMA) was selected to construct a biocompatible, field-responsive smart hydrogel material system. Adding different volumes of 1% methacrylated alginate (1:1, 1:2, 2:1) to the prepared polyacrylamide gel prepolymer solution yielded hydrogels with varying conductivity. The crosslinked hydrogels were then post-treated, including: to remove unreacted monomers, initiator fragments, and soluble oligomers, the prepared field-responsive smart hydrogel was immersed in deionized water, with the water changed every 12 hours for 24 hours. Next, to ensure sufficient ionization of the AlgMA units, the hydrogel was immersed in a 0.1 M NaOH solution for 1-2 hours to reach swelling equilibrium, followed by gentle rinsing with deionized water. Finally, the treated hydrogel was immersed in a 0.02 M NaCl solution for 24 hours to allow for complete equilibration. Before the experiment, the hydrogel was placed in DMEM for 2 hours to create a microenvironment suitable for cell / tissue growth.

[0040] like Figure 2As shown in the figure, this is a schematic diagram of a system used to quantify the cell's electrocoupling response. The hardware of this system can be considered as a chip. Figure A is an exploded view of the chip, including, from bottom to top: an adapter (blue), a PCB circuit board, a glass substrate, a chromium electrode, a gold electrode, a PDMS adhesive, a culture chamber, and a lid. The PCB circuit board and the glass gold electrode are connected by conductive silver paste. Figure B is a schematic diagram of fluorescence image acquisition and traction microscopy technology. The displacement and force fields of the fluorescent particles before and after deformation are calculated. Figure C shows the cell's active response (left): on a non-conductive hydrogel substrate, an electrical signal is applied to the cell, and the cell's electrical signal response is transmitted to the hydrogel deformation; passive response (right): on a conductive hydrogel substrate, electrical stimulation is applied to both the cell and the conductive hydrogel, and the hydrogel's electrical signal response is transmitted to the cell. Figures D and E show the synergistic loading of electrical signals: combining exogenous electrical signal loading with a stiffness-tunable hydrogel substrate.

[0041] The gold electrode glass chip in the cell-electric coupling chip system combines observation and electrical stimulation functions to acquire cell morphology and responses. The gold electrodes on the glass are fabricated using photolithography and magnetron sputtering techniques, where the photolithographically formed cutout pattern corresponds to the electrode pattern formed by magnetron sputtering. To ensure a tighter bond between the gold and glass, a layer of metallic chromium needs to be prepared before magnetron sputtering the gold. Based on the gold electrodes leading out from the glass substrate, a foldable printed circuit board (PCB) with four electrode leads was designed. The cutout in the middle perfectly matches the gold electrodes on the glass, and the electrodes around the glass are connected to the pads via conductive silver paste. The circuit is then connected to an external power source via a WAFER-GH1.25-4PLB connector.

[0042] To ensure sufficient cell flatness during observation and minimize the distance between cells and the confocal platform, thus avoiding image blurring in certain areas and exceeding the vertical imaging threshold during cell imaging, the thickness error of the gold electrode glass chip and PCB circuit board was measured to be approximately 0.1 mm before integration. Therefore, during integration, the PCB substrate needed to be thickened to match the height of the gold electrode glass chip, while simultaneously achieving a stable connection between the PCB pads and the gold electrode glass chip. Experimental results showed that using fluorescent particles as a target, the flatness of the glass substrate was sufficient for cell imaging. Most importantly, after the conductive silver paste connected the PCB circuit board to the gold electrodes on the glass, the resistance of all four electrodes in the circuit was approximately 4.5 Ω, thereby avoiding differences in cell dynamic response results caused by processing errors of different electrodes.

[0043] like Figure 3As shown in the figures, the fabrication process and characterization results of the glass-gold electrode chip are illustrated. Figure A shows the process flow of gold electrode fabrication using photolithography and magnetron sputtering techniques. Figure B shows the physical images of the gold electrode, including the image after photolithography (i) and the image after magnetron sputtering (ii). Figure C shows the design principle and signal transmission schematic diagram of the PCB circuit board. Figure D shows the thickness of the gold electrode glass and the PCB circuit board; statistics from 10 samples show that the PCB circuit board is approximately 0.95 mm thick, and the gold electrode glass is 1.05 mm thick. Figure E shows the assembly process of the cell-electric coupling chip, including the bonding of the glass-gold electrode to the glass ring, the UV fixation of the glass-gold electrode to the PCB circuit board, and the conductive connection. Figure F shows the assembled chip, with the resistance in the chip circuit after the PCB and glass-gold electrode are connected by conductive silver paste.

[0044] The mechanical properties of hydrogel substrates are crucial for characterizing cell dynamics. Substrate stiffness is an essential parameter in the inversion calculation of cell displacement and traction fields. Therefore, after preparing conductive and non-conductive hydrogels using different formulations, their stiffness was tested using atomic force microscopy (AFM). To ensure uniformity and generality of the tests, five equal regions were used, with 16 test points distributed in each region as a sample for testing results. Throughout the measurement process, the loading and unloading curves largely overlapped, and the slope of the test point during the sample contact phase represented the stiffness value. For non-conductive hydrogels, acrylamide and bisacrylamide monomers were used to regulate their stiffness. Based on this, a wide range of theoretical stiffness was used to prepare polyacrylamide gel prepolymers, and their stiffness was then tested. The experimental results showed that the average actual stiffness under different formulations were 0.75 kPa, 1.67 kPa, 12.87 kPa, 22.78 kPa, 37.51 kPa, and 41.17 kPa, respectively. The difference between theoretical and actual measured stiffness stems from the influence of materials and preparation environment. In the cell dynamics behavior inversion of this disclosure, the stiffness is calculated based on the actual measured value as input.

[0045] Based on this, the stiffness of the conductive hydrogel is also directly related to the deformation calculation under the action of an electrical signal. In this embodiment, the ion-conducting material selected is sodium methacrylamide (SMA). As it is a natural polymer crosslinking enhancer, its molecular chains can form an interpenetrating network (IPN) with the polyacrylamide (PAAM) in the non-conductive hydrogel through free radical polymerization. Simultaneously, the carboxyl and hydroxyl groups of sodium alginate can form hydrogen bonds and ion crosslinks, thus significantly improving the network density and mechanical strength of the conductive hydrogel system. Based on the above discussion, this embodiment selects three non-conductive hydrogel stiffnesses (1 kPa, 10 kPa, and 20 kPa) as the initial stiffness values ​​of the conductive hydrogel, and studies the response law between different contents of conductive material and electrical stimulation according to PAAm:AlgMA ratios of 1:1, 1:2, and 2:1, respectively. Using the same testing method, the results showed that, at an initial theoretical value of 1 kPa, the actual stiffness values ​​of gels with different proportions were approximately 14.52 kPa, 30.01 kPa, and 10 kPa, respectively. This trend also existed at initial theoretical values ​​of 10 kPa and 20 kPa. As mentioned earlier, the addition of AlgMA increased the stiffness of the system. The actual stiffness value of this conductive hydrogel provides a true reference for the inversion of electrically mediated hydrogel deformation, more accurately reflecting the dynamic-electric coupling response of cells.

[0046] The conductivity of conductive hydrogels also determines their deformation; therefore, the electrical properties of hydrogels require actual measurement results. To this end, the prepared conductive hydrogels were cut to known sizes, and their resistance values ​​were measured. Experimental results showed that, with constant stiffness, a higher AlgMA content resulted in lower resistance, i.e., greater conductivity, which is related to the proportion of conductive particles in the system. However, with a fixed PAAm to AlgMA ratio, there was no significant difference in conductivity at different stiffnesses (1 kPa, 20 kPa). In summary, the theoretical deformation of conductive hydrogels is directly proportional to the concentration of mobile ions in the system and inversely proportional to the stiffness.

[0047] Similarly, PEDOT:PSS was selected as the electronically conductive material in this embodiment. By incorporating different proportions of PEDOT:PSS, electrically tunable hydrogel substrates were constructed. The content of PEDOT:PSS in the precursor solution has a significant impact on the conductivity, polymerization efficiency, and microscopic imaging quality of the hydrogel. Therefore, after optimization, 0.1% and 1% PEDOT:PSS concentrations were selected as the conductive components for preparing electrically tunable hydrogels. As a polymer complex dispersed in the PAAm network, PEDOT:PSS, due to its inherent mechanical properties and molecular interactions, can alter the stiffness of the PAAm-PEDOT:PSS hydrogel. Young's modulus measurements were performed on hydrogels containing 0.1% and 1% PEDOT:PSS, and the results showed that the incorporation of PEDOT:PSS increased the stiffness of the hydrogel. When the PEDOT:PSS content was 0.1%, the modulus increased from 12.87 kPa to 14.56 kPa; when the PEDOT:PSS content was 1%, the modulus further increased to 16.02 kPa. Furthermore, the conductivity of the hydrogel itself is an important parameter for studying the relationship between electrical signals and cellular traction forces. The conductivity of the precursor solution and the polymerized hydrogel were measured. The results showed that the conductivity of the precursor solution was about an order of magnitude higher than that of the corresponding polymerized hydrogel, which may be due to the restricted effective orientation of PEDOT:PSS in the cross-linked gel network. For hydrogels containing 0%, 0.1%, and 1% PEDOT:PSS, the conductivity measured after gelation was 0.55 × 10⁻⁶ kPa. -4 1.32×10 -4 and 2.19×10 -4 S / m increases proportionally with increasing PEDOT:PSS content.

[0048] like Figure 4As shown in the figure, this diagram illustrates the design and characterization of a cell hydrogel substrate with tunable electrical properties and stiffness. Figure A shows a schematic diagram of hydrogel sampling for AFM testing, containing five sampling points (orange squares), each sampling point consisting of 4×4 array points, with each point spaced 2.5 μm apart. Figure B shows representative curves for AFM testing, with the blue line representing the loading curve and the red line representing the retraction curve. Young's modulus is the slope of the loading curve during the contact process with the hydrogel. Figure C shows the theoretical reference values ​​corresponding to different ratios of polyacrylamide monomers (acrylamide and methylenebisacrylamide). Figure D shows the actual Young's modulus of the polyacrylamide gel. Figure E shows the actual Young's modulus of the ion-conducting hydrogel (PAAm-AlgMA) with different ratios. Figure F shows the conductivity of hydrogels with different ratios at 1 kPa. Figure G is a schematic diagram of the preparation process of the electrically tunable hydrogel, which is treated with DMSO (dimethyl sulfoxide) and Triton X-100 to enhance the conductivity of the hydrogel. The figures show photographs of PAAm-PEDOT:PSS hydrogel prepolymers with PEDOT contents of 0%, 0.1%, and 1%, with the color gradually darkening. Figure H shows the Young's modulus values ​​of the electrically tunable hydrogels at PEDOT:PSS contents of 0.1% and 1%. Figure I shows the conductivity of the hydrogels with different PEDOT:PSS contents.

[0049] Tight bonding between the hydrogel and the glass substrate, as well as between the cells and the hydrogel, is fundamental for accurately characterizing the dynamic response of cells, avoiding the influence of substrate drift during cell response due to loose bonding. In this disclosure, a common glass activation method is used to enable the acryloyloxy and methacryloxy groups to form covalent bonds through carbon-carbon single bonds (CC), carbon-oxygen single bonds (CO), and carbon-oxygen double bonds (C=O), forming polymer chains that tightly bond the hydrogel to the glass substrate. Simultaneously, after surface modification of the hydrogel using Sulfo-SANPAH, it is activated under ultraviolet light to generate a highly reactive nitrene intermediate. This nitrene can undergo non-specific insertion reactions with multiple sites on the collagen I molecule, such as CH bonds and amine groups, forming stable CN covalent bonds, thereby achieving firm fixation of collagen I on the hydrogel surface.

[0050] Based on this, we treated adherent cells with electrical stimulation and lysis to investigate the displacement and traction fields of the cell's electrocoupling response under different voltages and stiffnesses. The results showed that, under the same stiffness, voltage was directly proportional to the cell's dynamic response induced by electrical stimulation. The active traction force of cells increased from approximately 60 Pa to approximately 180 Pa at high voltage compared to low voltage, an increase of about three times. Further, on a hard substrate, we also found that the active traction force of cells increased by about three times at high voltage compared to low voltage, further validating the relationship between voltage and cell dynamic response. Most importantly, orthogonal experiments revealed that the initial cell adhesion force was approximately 30 Pa on a soft substrate and approximately 45 Pa on a hard substrate. This indicates that the initial cell adhesion force is independent of voltage and only proportional to the substrate stiffness, thus satisfying the cell's resistance to hardness. Furthermore, on different conductive substrates, under the same applied voltage and substrate stiffness, the experimental results showed that appropriate substrate conductivity can enhance cell traction, while excessively high conductivity weakens the traction force. Specifically, when the PEDOT:PSS content was 0.1%, the cell traction force was approximately 200 Pa; however, when the PEDOT:PSS content increased to 1%, the traction force decreased to approximately 70 Pa. This phenomenon is closely related to the current density at the cell-substrate interface obtained through finite element simulation in the previous section. On a non-conductive substrate, the surface current densities were 0.51 and 1.02 μA / cm² when applied at 2.5 V and 5 V, respectively. After introducing PEDOT:PSS, at 2.5 V, the current density increased to 1.151 μA / cm² with 0.1% PEDOT:PSS and to 2.093 μA / cm² with 1% PEDOT:PSS. These results indicate that optimal current stimulation can enhance cell-substrate adhesion and increase traction force, while excessive current stimulation leads to a decrease in traction force.

[0051] like Figure 5 As shown in the figure, the electrocoupling response of cells on electrically tunable and stiffness-tunable hydrogel substrates is characterized. Figure AB shows the inversion contour plots and traction force statistics of voltage versus cell active response on soft substrates. Figure CD shows the inversion results and statistics of voltage versus cell active response on hard substrates. Figure EF shows the displacement field and traction force field contour plots and statistics of cells on PEDOT:PSS conductive hydrogel.

[0052] Polyacrylamide (PAAm) and sodium methacrylamide prepolymer (AlgMA) form a three-dimensional network structure through free radical polymerization in the presence of a crosslinking agent and an initiator. The carboxyl group (-COOH) in AlgMA is a weak acid group, which ionizes to -COO under alkaline conditions. -This process imparts a negative charge to the entire hydrogel network, resulting in a polyanionic electrolyte gel. When placed in an electrolyte solution (such as NaCl solution) and a direct current electric field is applied, the anions (-COO-) inside the gel... - ) remains fixed, while cations (such as Na) + The migration of ions towards the cathode creates an ion concentration and osmotic pressure difference across the gel. According to the principle of osmosis, water molecules migrate from areas of low ion concentration (low osmotic pressure) to areas of high ion concentration (high osmotic pressure), causing the cathode side to absorb water and swell, while the anode side dehydrates and shrinks. Ultimately, this leads to the gel bending towards the anode.

[0053] Generally, electrode shape is closely related to the deformation effect of hydrogel, and a suitable electrode shape design can more easily reflect the influence between its shape and deformation. Simultaneously, electric field parameters also affect the deformation response; minute changes caused by excessively small electric field conditions may be difficult to capture, while large deformations caused by excessively large electric field conditions will exceed the image acquisition area, severely affecting the inversion calculation results. Based on this, this disclosure first uses finite element simulation to evaluate the potential and electric field distributions in electrically mediated hydrogels under two electrode shapes (square and circular). Simulation results show that when square and circular electrodes are used as positive and negative electrodes, the potential distribution around the square electrode is more uniform, while the potential distribution around the circular electrode varies significantly. Similar results were found in the electric field results. Importantly, the electric field contour plot reveals that due to the sharp-point effect generated by the corners of the square electrode, a higher electric field appears at the top and bottom corners of the square electrode, which then diffuses and decreases towards the surrounding area.

[0054] Building upon this, this disclosure investigates the relationship between different electrode shapes and electroinduced hydrogel deformation. A square electrode was used as the negative electrode, and a circular electrode as the positive electrode. Based on previous theoretical and experimental results, the phenomena of positive electrode contraction and negative electrode swelling were observed. First, fluorescence microscopy images were acquired at different time points. Inversion calculations showed that the hydrogel deformation of both the positive and negative electrodes gradually increased with electrical stimulation time, eventually stabilizing. Most importantly, the deformation at the positive electrode (circular electrode) was clearly arc-shaped, while that at the square electrode was smoother, especially at the 5th second, where a significant deformation at both ends due to the tip effect was clearly visible.

[0055] Furthermore, on a time scale, it was found that the electrodeformable hydrogel exhibited greater changes in the early stages, with the changes gradually decreasing over time. The X and Y positions of the maximum values ​​in the cloud maps at 0 / 1 / 5 / 10 / 15 / 20 seconds were statistically analyzed, and the results were consistent with the previous findings.

[0056] Electrodeformable ion-conductive hydrogels are not only affected by electrode shape, but their deformation behavior is also closely related to electric field parameters and ion content. Based on this, this disclosure first increases the input voltage to investigate the relationship between input voltage and hydrogel deformation. The results show that doubling the input voltage increases the deformation of the conductive hydrogel at the same time point by a corresponding amplitude. Preliminary results indicate a positive correlation between input voltage and conductive hydrogel deformation. That is, increasing the input voltage can reduce the time to reach the target deformation. The ion concentration in the electrodeformable hydrogel system is a crucial factor determining the deformation; however, an appropriate content is needed to balance the relationship between hydrogel properties and the final deformation. As the above theory and results show, polyacrylamide gel possesses good mechanical properties, while sodium methacrylamide alginate possesses good electrical conductivity. Therefore, an appropriate ratio is necessary to better leverage the advantages of both materials, ensuring that the electrodeformable hydrogel possesses both excellent mechanical and deformation properties. Based on the above discussion, this embodiment further explores the effects of low (PAAm:AlgMA 2:1) / high (PAAm:AlgMA 1:2) ion concentrations on hydrogel deformation under the original ratio (PAAm:AlgMA 1:1). Experimental results show that the higher the AlgMA content in the system, the greater the deformation of the hydrogel under the same electrical parameters. Quantitative calculations using inversion show that the hydrogel deformation caused by the two ion concentrations is approximately 6 times, a trend consistent with osmotic pressure theory. Statistical results clearly confirm this pattern, especially the comparison between the two ion concentration systems, which more clearly demonstrates the direct relationship between ion concentration and deformation in the conductive hydrogel system.

[0057] like Figure 6 Figure 1 shows the characterization of the electrocoupling response of the ion-conductive hydrogel. Figure A illustrates the electrical signal loading method and image acquisition area of ​​the ion-conductive hydrogel. Figure B shows the potential and electric field cloud maps from the finite element simulation. Figure C shows the displacement field distribution obtained from the fluorescent particle images acquired through TFM inversion. Figures DE show the results of statistical analysis of the images of the anode (D) and cathode (E) at the 20th second. Figure F shows the longitudinal (top) and transverse (bottom) data acquisition and analysis at the maximum displacement of the anode and cathode. Figures GH show the displacement statistics at representative time points.

[0058] The preparation method of the gold electrode may include: Gold electrode glass chips were fabricated using photolithography and etching techniques. Electrode patterns were designed using CAXA drawing software, and film masks for the gold electrode glass were obtained using inkjet direct writing technology. First, the glass (40×40 mm, 1 mm thick) was cleaned by ultrasonic cleaning three times each with acetone, anhydrous ethanol, and deionized water for 5 minutes each time, followed by drying with nitrogen. SU8-2050 negative photoresist was spin-coated onto the glass surface. Based on experience, a 75 μm thickness was achieved, requiring pre-coating at 500 rpm for 25 seconds and spin-coating at 2200 rpm for 50 seconds. The spin-coated glass was then pre-baked on hot plates at 65 ℃ and 95 ℃ for 2.5 minutes and 8.5 minutes, respectively, to reduce thermal stress on the photoresist layer. The glass was then removed from the hot plates and cooled to room temperature. The spin-coated photoresist was then exposed to ultraviolet light through a film mask with a circuit diagram designed on it using an exposure machine. The measured power P (mW / cm²) was used to determine the optimal exposure time. 2 ) and the required exposure W (mJ / cm 2 The exposure time was calculated. The exposed glass was placed on hot plates at 65 ℃ and 90 ℃ for 1.5 minutes and 6.5 minutes respectively, then removed from the hot plates and cooled to room temperature. The exposed glass was placed in a glass dish, and an appropriate amount of developer (SU8 developer) was poured in and manually and slowly applied until the pattern at the exposed area was fully visible. Immediately afterwards, it was rinsed with isopropanol to remove residual resist, then rinsed with deionized water on both sides and dried with nitrogen. To improve the adhesion between the resist layer and the substrate, the cleaned chip was placed on a hot plate at 150 ℃ for 30 minutes to harden the film. The photolithographically etched glass was then magnetron sputtered (using an ultra-high vacuum magnetron sputtering system, TEC-SPU-400-1) to apply an adhesion layer (chromium, Cr, 3-5 nm) and an electrode layer (gold, Au, 100 nm). Finally, the gold-sputtered glass was completely immersed in a resist remover solution for ultrasonic removal until the gold electrode was fully visible on the glass surface.

[0059] The design of PCB circuit boards and the assembly of cell culture dishes may include: The cell culture device consists of a bottomless cell culture plate, patterned gold electrode glass, and a PCB circuit board.

[0060] Assembly of the glass ring: First, clean the glass electrode and glass ring with 75% alcohol, dry with nitrogen, rinse with deionized water, and place in a 50 ℃ forced-air drying oven until completely dry. Using a prepolymer of polydimethylsiloxane (PDMS, Sylgard 184, Dow Corning) at a weight ratio of 10:1 (base / curing agent), the bottomless cell culture plate was adhered to the prepared gold electrode glass and dried at 65 ℃ for 2 hours.

[0061] The design and assembly of PCB circuit boards may include: To ensure the stability of the electrical response experiment, electrodes were led out and fixed using a PCB circuit board. First, a four-electrode PCB circuit board was designed, with a central cutout for mounting glass. The electrodes around the glass were connected to the pads using conductive silver paste, and connected to an external power supply via a WAFER-GH1.25-4PLB connector. Specifically, a square pad component and its package were first created, with the package set as the top-level pad. Then, the WAFER-GH1.25-4PLB connector was connected to this pad component. According to the schematic, routing was completed on the top layer with a line width of 10 mil (0.254 mm), reliably connecting the pad to the connector. Finally, FR-4 substrate was selected, and the electrode portions were immersion gold treated. During assembly, considering the thickness difference between the PCB board (~0.95 mm) and the glass (~1.05 mm), and to prevent the gold electrode glass from exceeding the microscope's sampling threshold after being suspended in the air, a standard A4 paper (~0.1 mm) was first applied to the bottom layer of the PCB. Then, the PCB board and the gold electrode glass were assembled, and weights were used to press down the glass ring, ensuring complete contact between the gold electrode glass and the bottom surface. UV-curing adhesive (ergo8500) was used to quickly fix the glass and PCB (UV curing for 60 seconds). Finally, conductive silver paste was used to connect the four corresponding contacts of the gold electrode glass to the PCB board, and the assembly was cured in a 180 ℃ oven for 60 minutes.

[0062] Cytotoxicity of conductive / non-conductive hydrogels: The cells used in this disclosure were mouse intestinal epithelial cells (IEC-6), cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (PS) in a cell culture incubator at 37 °C and 5% CO2. The solidified hydrogel sample, after thorough washing to remove monomers, was placed in a cell culture plate. The hydrogel was then treated with sulfo-SANPAH and incubated with collagen (Collagen I). Finally, a digested IEC-6 cell suspension was uniformly seeded onto the hydrogel surface, and the culture plate was returned to the incubator for 24 hours. Staining was performed using Calcein AM (green fluorescence for live cells), Propidium Iodide (red fluorescence for dead cells), and Hoechst 33342 (nuclear dye for live cells). Images were acquired using a fluorescence microscope, and the ratio of live to dead cells was quantitatively analyzed using ImageJ software.

[0063] Young's modulus test of hydrogels: The mechanical properties of hydrogels are key characteristics affecting their deformation. The Young's modulus of hydrogels was measured using a bio-Atomic Force Microscope (BioScope Resolve). The experiment employed PeakForce QNM (Quantitative Nanomechanical Mapping) in a liquid environment, with a Hertzian (Spherical) contact mechanics model. A silicon nitride probe (model: MLCT-SPH-1UM) with a tip radius of 970 nm was used. The calibrated probe parameters were: sensitivity 70.79 nm / V, spring constant 0.02800 N / m, and Poisson's ratio set to 0.5. During the measurement, the probe operated at a speed of 16.1 μm / s, scanning five regions of the sample using a 4×4 matrix with a dot spacing of 2.5 μm and a 1 mm interval between each matrix. The collected data were processed using AFM analysis software (NanoScope Analysis 1.8).

[0064] Inversion of displacement and traction fields: The cellular traction force field and displacement field are obtained through traction force microscopy (TFM). By identifying and locating fluorescent particles, the electrical response displacement field of the conductive substrate can be obtained. Simultaneously, combined with the elastic modulus of the substrate, the active traction force field of the cell is obtained through inversion, i.e., TFM inversion. Specifically, fluorescent particles (beads) are uniformly distributed on the gel surface as tracer particles. After cells are in close contact with the hydrogel surface through collagen I, an electrical signal is applied to the non-conductive polyacrylamide gel platform, causing a cellular response. The cells, in response, deform the hydrogel, resulting in displacement changes of the fluorescent particles inside the hydrogel. A series of images before and after the deformation of the fluorescent particles are acquired using a confocal microscope. Inversion parameters, sampling parameters, and image parameters are set. Inversion parameters include the true stiffness and thickness of the hydrogel. Sampling parameters include a confocal sampling mode, an exposure time of 1 second, an excitation wavelength of 561.5 nm, and an emission wavelength of 595 nm. Image parameters include pixel size, sampling duration, a numerical aperture of 0.75, and a camera depth of 8 bits. In a series of images, a local area, i.e., the rigid body translation calculation region, is selected. This selected region is input into an inversion program, such as MATLAB, where the deformation field, i.e., the displacement field, is obtained using digital image correlation methods. Furthermore, inversion parameters, sampling parameters, and image parameters are input into the inversion program. The program then uses the displacement field and the input parameters to invert the traction force field. Specifically, the actual stiffness of the hydrogel used can be 1 kPa, 10.61 kPa, or 19.66 kPa, and the hydrogel thickness can be 100 μm. The regularization parameter can be set to 0.0001 by default.

[0065] This disclosure provides an electrical coupling method for studying single-cell-substrate interactions. The method employs the electrical coupling chip system described in any of the above methods, and the method includes generating cell dynamic behavior information through the electrical coupling chip system.

[0066] This disclosure provides an electrical coupling device for studying single-cell-substrate interactions. The device is used to perform an electrical coupling method for studying single-cell-substrate interactions as described in the above embodiments. The device includes a generation module configured to generate kinetic behavior information of the cell through the cell electrical coupling chip system.

[0067] The cell power coupling device and the cell power coupling method provided in the above embodiments of this disclosure are based on the same inventive concept and have the same beneficial effects as the methods used, run or implemented by the applications stored therein.

[0068] This disclosure also provides an electronic device corresponding to the cell-electric coupling method provided in the foregoing embodiments, for executing the aforementioned electric coupling chip system for studying single-cell-substrate interactions. This disclosure is not limiting.

[0069] It should be noted that: In the foregoing text, the terms "comprising," "including," or any other variations thereof are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element. Furthermore, it should be noted that the scope of the methods and apparatuses in the embodiments of this disclosure is not limited to performing functions in the order shown or discussed, but may also include performing functions substantially simultaneously or in the reverse order, depending on the functions involved. For example, the described methods may be performed in a different order than described, and various steps may be added, omitted, or combined. Additionally, features described with reference to certain examples may be combined in other examples.

[0070] Through the above description of the embodiments, those skilled in the art can clearly understand that the methods of the above embodiments can be implemented by means of software plus necessary general-purpose hardware platforms. Of course, they can also be implemented by hardware, but in many cases the former is a better implementation method. Based on this understanding, the technical solution of this disclosure, in essence, or the part that contributes to the prior art, can be embodied in the form of a software product. This computer software product is stored in a storage medium (such as ROM / RAM, magnetic disk, optical disk), and includes several instructions to cause a terminal (which may be a mobile phone, computer, server, air conditioner, or network device, etc.) to execute the methods described in the various embodiments of this disclosure.

[0071] The embodiments of this disclosure have been described above with reference to the accompanying drawings. These are merely specific implementations of this disclosure, but this disclosure is not limited to the specific implementations described above. The specific implementations described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of this disclosure without departing from the spirit and scope of the claims, and all of these forms are within the protection scope of this disclosure.

Claims

1. A power-coupled chip system for studying single-cell-substrate interactions, characterized in that, The system includes a first subsystem and a second subsystem. Each subsystem includes a culture module, an observation module, and an electrical signal transmission module. The system also includes a post-processing module. The culture module includes a cell culture chamber comprising a transparent glass and a hydrogel placed on the transparent glass. The hydrogel is used to support and culture cells. The hydrogel in the first subsystem is a conductive hydrogel, and the hydrogel in the second subsystem is a non-conductive hydrogel. The conductive hydrogel includes electronic conductivity and ionic conductivity. By changing the electrical and mechanical properties of the hydrogel, the cells can be made to respond passively. The cells supported by the non-conductive hydrogel are used to directly receive electrical and mechanical signals to obtain an active cell response. The electrical signal transmission module includes a PCB circuit board for transmitting electrical stimulation signals and transmission electrodes; The observation module includes an imaging device that captures images of the culture chamber during the electrical response process through the transparent glass to obtain images; The post-processing module is used to post-process the images obtained by each subsystem to obtain information on the dynamic behavior of cells; Fluorescent particles exist on the surface of the hydrogel. Under the action of electrical coupling signals, the cell response is transmitted to the hydrogel substrate and expressed through the fluorescent particles. The post-processing module is used to perform inversion calculations by analyzing the positions of fluorescent particles in the images acquired by each subsystem.

2. The system according to claim 1, characterized in that, The images include images before deformation and images after deformation, the imaging device is a high-speed fluorescence microscope, and the dynamic behavior information includes displacement field and traction force field; The high-speed fluorescence microscope is used to capture images of cells in real time before the electrocoupling process to obtain images before deformation; and to capture images of cells in real time after the electrocoupling process to obtain images after deformation. The post-processing module is used to perform inversion calculations on the captured images to obtain the displacement field and traction field of the cell under the action of the electrical coupling signal.

3. The system according to claim 1, characterized in that, The PCB circuit board includes PCB pads, adapters, and PCB electrodes; The transfer electrode includes a chromium electrode and a gold electrode. The chromium electrode is disposed on the upper surface of the transparent glass, and the gold electrode is disposed above the chromium electrode. A closed loop between the electrodes is formed in the culture chamber, the gold electrode, and the PCB electrode. The gold electrode includes a symmetrical electrode and an asymmetrical electrode. The symmetrical electrode is used to apply electrical stimulation with a uniform electric field, and the asymmetrical electrode is used to apply electrical stimulation with a non-uniform electric field.

4. The system according to claim 1, characterized in that, The system also includes an electrode generation module; The electrode generation module is used to generate the gold electrode in the transport electrode by photolithography and magnetron sputtering techniques, according to the set electrode size, electrode shape, array size and array density.

5. The system according to claim 3, characterized in that, The gold electrode shape includes square and round; The PCB circuit board is a square printed circuit board. The PCB pads of the PCB circuit board are provided with multiple electrode leads. The hollow shape and thickness of the inner ring of the PCB circuit board are consistent with the outer contour shape and thickness of the gold electrode. The gold electrodes located around the transparent glass are connected to the PCB pads via conductive silver paste, and the adapter is used to receive external electrical signals.

6. The system according to claim 1, characterized in that, The ion concentration, stiffness, and voltage of the electrical stimulation signal of the hydrogel are preset variables, which are related to the magnitude of the cell response of the hydrogel during the electrical coupling process.

7. The system according to claim 1, characterized in that, The conductive hydrogel is an acrylamide and sodium methacrylamide / PEDOT:PSS gel, wherein the combination of acrylamide and PEDOT:PSS constitutes an electronically conductive hydrogel, and the combination of acrylamide and sodium methacrylamide constitutes an ionically conductive hydrogel, and the non-conductive hydrogel is a polyacrylamide gel.

8. A method for generating cell information, characterized in that, The method employs a power-coupled chip system for studying single-cell-substrate interactions according to any one of claims 1-7, the method comprising: The power-coupled chip system generates information about the dynamic behavior of cells.

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