Multi-flux real-time electric monitoring heart chip

Abstract

The utility model belongs to the technical field of heart chips, and particularly relates to a multi-flux real-time electric monitoring heart chip which comprises a perfusion layer, a culture room layer and a glass substrate-electrode plate which are fixedly connected in sequence from top to bottom, a tree-shaped step perfusion channel is formed in the perfusion layer, the tree-shaped step perfusion channel comprises n stages of mixing flow channel groups which are sequentially arranged in a communicating mode in the liquid flowing direction, the liquid outlet end of the mixing flow channel group located at the tail end communicates with the liquid inlet end of a liquid injection part, and the liquid outlet end of the liquid injection part communicates with an outlet; the culture room layer is provided with a plurality of culture parts, the culture parts are in one-to-one correspondence with the liquid injection parts, and the liquid inlet ends of the culture parts are communicated with the middle parts of the liquid injection parts; a plurality of electrode parts are arranged on the glass substrate-electrode plate, and the plurality of electrode parts and the plurality of culture parts are in one-to-one correspondence and are connected.

Description

Technical Field

[0001] The utility model belongs to the technical field of heart chips, and in particular relates to a multi-flux real-time electrical monitoring heart chip. Background Art

[0002] Heart disease is a significant burden on global health care systems and the leading cause of death each year. To improve our understanding of heart disease, high-quality disease models and efficient drug screening methods are needed. The rapid development of microfluidics and soft lithography combined with advances in human induced pluripotent stem cell (iPSC) biology has led to a new field of "organs on a chip" that aims to build controllable, minimum functional units that can reproduce specific organ-level functions in vitro.

[0003] Combined with microfluidics technology, long-term dynamic culture and observation of cells in a closed environment can be achieved; combined with sensing technology (electrical sensing, pressure sensing) and biomaterials: poly(maleic acid octamethylene (anhydride) citrate (POMaC) and crack sensor integrated cantilever can monitor changes in myocardial tissue contractility, gold electrodes and platinum wire electrodes can be used to monitor electrical signals, and inverse opal hydrogel and capillaries can be used to visualize myocardial contraction. Although the construction of the heart chip platform has a solid foundation, the contradiction between bionics and flux integration still needs to be resolved, and efficient drug screening functions cannot be achieved on a single platform.

[0004] In response to the problems of low flux and insufficient bionics in current heart chips, we propose a multi-flux real-time electrical monitoring heart chip. Utility Model Content

[0005] The utility model aims to provide a multi-flux real-time electrical monitoring heart chip to solve the above problems.

[0006] To achieve the above purpose, the utility model provides the following solutions:

[0007] A multi-flux real-time electrical monitoring heart chip, comprising: a perfusion layer, a culture chamber layer and a glass substrate-electrode plate fixed in sequence from top to bottom;

[0008] The perfusion layer is provided with a tree-like stepped perfusion channel, and the tree-like stepped perfusion channel includes n-level mixing channel groups that are sequentially connected and arranged along the liquid flow direction, where n is a positive integer, and the mixing channel group includes n+1 liquid inlet ends and n+2 liquid outlet ends; the liquid outlet end of the previous mixing channel group corresponds to and is connected to the liquid inlet end of the next mixing channel group;

[0009] The liquid outlet ends of the mixing flow channel group located at the tail end are respectively connected to the liquid inlet ends of the liquid injection part, and the liquid outlet ends of the liquid injection part are connected to the outlet;

[0010] A plurality of culture parts are provided on the culture chamber layer, the plurality of culture parts correspond to the plurality of liquid injection parts one by one, and the liquid inlet end of the culture part is connected to the middle of the liquid injection part;

[0011] The glass substrate-electrode plate is provided with a plurality of electrode parts, and the plurality of electrode parts correspond to and are connected with the plurality of culture parts one by one.

[0012] Preferably, the liquid injection portion comprises a plurality of perfusion chambers connected in sequence, the liquid inlet end of the perfusion chamber located at the front end in the liquid flow direction is connected to the liquid outlet end of the corresponding mixing channel group, and the liquid outlet end of the perfusion chamber located at the rear end in the liquid flow direction is connected to the corresponding outlet;

[0013] A plurality of the perfusion chambers are communicated with the corresponding culture parts.

[0014] Preferably, the culture part comprises a plurality of culture chambers, the plurality of culture chambers correspond one-to-one to the plurality of perfusion cavities, the culture chambers are connected to the perfusion cavities, and one end of the electrode part is correspondingly arranged in the culture chamber.

[0015] Preferably, the electrode portion includes two internal signal collection targets, the internal signal collection targets are electrically connected to edge output signal targets, and the internal signal collection targets are fixed to the bottom of the corresponding culture chamber.

[0016] Preferably, the liquid injection part includes three perfusion chambers, the three perfusion chambers are connected in sequence, the perfusion chamber is an elliptical structure of 3.5mm*6mm, and the height of the perfusion chamber is 150um.

[0017] Preferably, the culture part includes three culture chambers, each of which is an elliptical structure of 3.5 mm*6 mm, and has a thickness of 2 mm.

[0018] A method for manufacturing a multi-flux real-time electrical monitoring heart chip, based on the above multi-flux real-time electrical monitoring heart chip, comprises the following steps:

[0019] S1. respectively preparing molds having the perfusion layer structure and the culture chamber layer structure;

[0020] S2, injecting a mixture of PDMS prepolymer A and crosslinking agent B into the mold and curing it to obtain the perfusion layer and the culture chamber layer respectively, and then cleaning and sterilizing them;

[0021] S3, drilling a hole at a designated position in the culture part of the culture chamber layer, bonding the culture chamber layer to the glass substrate-electrode plate, and then cleaning and sterilizing;

[0022] S4. After inoculating cells in the culture chamber layer, place the layer in an incubator. After the cells adhere to the wall, bond the perfusion layer to the culture chamber layer to form a complete chip system.

[0023] Preferably, in step S1, the method for manufacturing the mold comprises the following steps:

[0024] S101, spin coating SU-8 photoresist on the silicon wafer, pre-baking at 95° for 30 min;

[0025] S102, placing the mask on the silicon wafer and then UV-exposing it, followed by post-baking at 95° for 30 minutes;

[0026] S103, using ethyl lactate for development, hardening the film, temperature 120°, time 30 min.

[0027] Preferably, in step S2, the PDMS prepolymer A and the cross-linking agent B are mixed in a ratio of 10:1, the mixed solution is injected into a mold, and then vacuum degassed. After degassed, the mixture is cured in an oven at 80° C. for 1 hour to obtain the perfusion layer and the culture chamber layer.

[0028] Compared with the prior art, the utility model has the following advantages and technical effects:

[0029] When in use, the injection pump is connected to the liquid inlet of the tree-like stepped perfusion channel through a PTFE tube, a flat needle and a syringe. There are two liquid inlet ends of the mixing flow channel group at the front end of the liquid inflow direction of the tree-like stepped perfusion channel, one of which is used to inject a culture medium with a specific concentration of drugs, and the other is used to inject a drug-free culture medium. When the culture medium with a specific concentration of drugs and the drug-free culture medium are injected at the same time, a drug concentration gradient is automatically formed under the action of the tree-like stepped perfusion channel, and different concentrations of drug solutions enter the corresponding liquid injection part, and are injected from the liquid injection part into the corresponding culture part opened on the culture chamber layer. Myocardial cells are cultured in the culture part. When the drug solution fills the culture part, it will flow out through the outlet. The electrode parts arranged in each culture part are used to electrically stimulate the myocardial cells and detect the cell field potential. The state of the myocardial cells can be detected under different drug solution concentrations. The utility model can realize the co-culture of heterotypic cells in the same system and be used in cell interaction research. The microelectrode array design significantly improves the real-time electrophysiological evaluation flux compared with the existing heart chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the embodiments of the utility model or the technical solutions in the prior art, the drawings required for use in the embodiments are briefly introduced below. Obviously, the drawings described below are only some embodiments of the utility model. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor:

[0031] Figure 1 This is a schematic diagram of the structure of the perfusion layer of the utility model;

[0032] Figure 2 This is a schematic diagram of the layer structure of the culture chamber of the utility model;

[0033] Figure 3 This is a schematic diagram of the glass substrate-electrode plate structure of the utility model;

[0034] Figure 4 This is a schematic diagram of the relative position of the internal signal collection target and the culture chamber of the utility model;

[0035] Figure 5 This is the concentration gradient distribution diagram of the utility model;

[0036] Figure 6 This is the outlet concentration distribution diagram of the utility model;

[0037] Figure 7 It is a schematic diagram of the structure of the utility model;

[0038] Figure 8 Make a process flow chart for this utility model;

[0039] Fig. 9 This is a schematic diagram of the use state of the utility model;

[0040] Fig.10 This is the waveform diagram of the in situ myocardial cell field potential acquisition of the utility model;

[0041] Fig.11 Characterization of the concentration screening function of this utility model - detection of secretory proteins from cardiomyocytes;

[0042] Among them, 1. perfusion layer; 2. culture chamber layer; 3. glass substrate-electrode plate; 4. tree-like stepped perfusion channel; 5. perfusion cavity; 6. culture chamber; 7. internal signal collection target; 8. edge output signal target; 9. exit. DETAILED DESCRIPTION

[0043] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0044] In order to make the above-mentioned purposes, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0045] Reference Figures 1 to 11 The utility model discloses a multi-flux real-time electrical monitoring heart chip, comprising: a perfusion layer 1, a culture chamber layer 2 and a glass substrate-electrode plate 3 fixedly connected in sequence from top to bottom;

[0046] A tree-like stepped perfusion channel 4 is provided on the perfusion layer 1. The tree-like stepped perfusion channel 4 includes n-level mixing channel groups that are sequentially connected and arranged along the liquid flow direction. n is a positive integer. The mixing channel group includes n+1 liquid inlet ends and n+2 liquid outlet ends. The liquid outlet end of the previous mixing channel group corresponds to and is connected to the liquid inlet end of the next mixing channel group.

[0047] The liquid outlet ends of the mixing flow channel group at the tail end are respectively connected to the liquid inlet ends of the liquid injection part, and the liquid outlet ends of the liquid injection part are connected to the outlet 9;

[0048] A plurality of culture parts are provided on the culture chamber layer 2, and the plurality of culture parts correspond to the plurality of liquid injection parts one by one, and the liquid inlet end of the culture part is connected to the middle of the liquid injection part;

[0049] A plurality of electrode parts are arranged on the glass substrate-electrode plate 3, and the plurality of electrode parts correspond to and are connected with the plurality of culture parts one by one.

[0050] When in use, the injection pump is connected to the liquid inlet of the tree-like step perfusion channel 4 through a PTFE tube, a flat needle and a syringe. There are two liquid inlet ends of the mixing channel group at the front end of the liquid inflow direction of the tree-like step perfusion channel 4, one of which is used to inject a culture medium with a specific concentration of drugs, and the other is used to inject a drug-free culture medium. When the culture medium with a specific concentration of drugs and the drug-free culture medium are injected at the same time, under the action of the tree-like step perfusion channel 4, a drug concentration gradient is automatically formed, and different concentrations of drug solutions enter the corresponding liquid injection part, and are injected from the liquid injection part into the corresponding culture part opened on the culture chamber layer 2. Myocardial cells are cultured in the culture part. When the drug solution fills the culture part, it will flow out through the outlet 9. The electrode parts arranged in each culture part are used to electrically stimulate the myocardial cells and detect the cell field potential. The state of the myocardial cells can be detected at different drug solution concentrations. The utility model can realize the co-culture of heterotypic cells in the same system and be used in cell interaction research. The microelectrode array design significantly improves the real-time electrophysiological evaluation flux compared with the existing heart chip.

[0051] In a further optimized solution, the liquid injection portion includes a plurality of perfusion chambers 5 connected in sequence, the liquid inlet end of the perfusion chamber 5 located at the front end of the liquid flow direction is connected to the liquid outlet end of the corresponding mixing channel group, and the liquid outlet end of the perfusion chamber 5 located at the rear end of the liquid flow direction is connected to the corresponding outlet 9;

[0052] A plurality of perfusion chambers 5 are communicated with corresponding culture parts.

[0053] According to a further optimized solution, the culture part includes a plurality of culture chambers 6 , which correspond one to one with a plurality of perfusion chambers 5 , the culture chambers 6 are connected with the perfusion chambers 5 , and one end of the electrode part is correspondingly arranged in the culture chambers 6 .

[0054] According to a further optimized solution, the electrode portion includes two internal signal collection targets 7 , the internal signal collection targets 7 are electrically connected to the edge output signal targets 8 , and the internal signal collection targets 7 are fixed at the bottom of the corresponding culture chamber 6 .

[0055] In view of the current problems of low flux and insufficient bionics in heart chips, the utility model constructs a microfluidic real-time dynamic monitoring heart chip platform, in order to realize the integration of flux and bionic mechanism technology, and apply it to the research of heart disease modeling, drug screening, cell interaction, etc., to explore the laws and mechanisms of disease occurrence and development, and to find disease markers and drug targets, providing new tools for related biomedical research.

[0056] The utility model is a layer-by-layer stacking structure, which is divided into three parts. The top layer is a perfusion layer 1, and a tree-like stepped perfusion channel 4 and a plurality of perfusion chambers 5 are arranged on the perfusion layer 1: the perfusion chamber 5 of the culture chamber is an elliptical structure of 3.5mm*6mm, and a plurality of perfusion chambers 5 are distributed in an array. The height of the perfusion chamber 5 is preferably 150um. The utility model preferably has 30 perfusion chambers 5, which are distributed in 10 rows and 3 columns; the perfusion layer 1 has a thickness of 2mm, and the tree-like stepped perfusion channel 4 has a height of 150um. The bottom surface of the perfusion layer 1 is in direct contact with the top surface of the culture chamber layer 2, and each perfusion chamber 5 is docked and connected with the top of the corresponding culture chamber 6; there are two liquid inlet ends of the mixing flow channel group located at the front end of the liquid inflow direction of the tree-like stepped perfusion channel 4, and the diameter of the outlet 9 is 1mm. The utility model preferably has 10 outlets 9. After the culture medium containing a specific concentration of the drug and the culture medium without the drug are respectively perfused from the inlet, the outlet 9 can obtain a linear concentration gradient culture medium containing the drug for forming different environments for cell culture.

[0057] refer to Figures 4 to 5 , the layer can form a stable linear concentration gradient through simulation by comsolmultiphysisc software: the concentration at the right outlet is linear from top to bottom, and the simulated data is plotted to obtain Figure 5 ,Depend on Figure 5 It can be seen that the outlet concentration is linearly related.

[0058] The middle layer is the culture chamber layer 2: a number of culture chambers 6 are provided on the culture chamber layer 2, and the culture chambers 6 are 3.5mm*6mm elliptical structures, and the number of culture chambers 6 are distributed in an array. The utility model preferably has 30 culture chambers 6, and the culture chambers 6 are arranged one by one with the perfusion chambers 5. The culture chambers 6 are 2mm thick, and the wide diameter of the culture chambers 6 is 3.5mm, and the long diameter is 6mm. This layer is used to inoculate cells for cell culture. The culture chambers 6 are connected to the corresponding perfusion chambers 5 on the top, and the culture chambers 6 are in contact with the glass substrate-electrode plate 3 on the bottom. The independent perfusion chambers 5 and culture chambers 6 allow the culture medium of the perfusion layer 1 to be infiltratively filled with the culture chambers 6, so as to reduce the impact force of the perfusion on the adherent cells.

[0059] The glass substrate-electrode plate 3 has a thickness of 1 mm. On its top surface (the surface in contact with the culture chamber layer 2), titanium is first plated with a thickness of 20 nm, and then gold is plated with a thickness of 50 nm to form a microelectrode array: the internal signal collection target 7 is a 0.4 mm × 0.4 mm rectangle, and the edge output signal target 8 is a 1.37 mm × 1.37 mm rectangle, and the two are connected by a 200 um wide passage. Two internal signal collection targets 7 are set at the bottom of each culture chamber 6.

[0060] The microelectrode array can perform real-time electrical stimulation and field potential acquisition during the cell culture process: the signal generator is connected to the output signal target, and the stimulation signal is transmitted to the internal acquisition signal target 7 through the microelectrode array to act on the cells, thereby controlling the cell beating frequency and amplitude; conversely, the microelectrode array can transmit the field potential generated by the cell beating to the edge output signal target 8, and the edge output signal target 8 is connected to the signal amplifier to convert the cell field potential into a visual waveform.

[0061] This platform is more versatile than the current heart chip, and can realize efficient drug screening functions on a single platform, realize the co-culture of heterotypic cells in a closed system, and improve the throughput of real-time electrophysiological evaluation.

[0062] Multi-flux: With the increasing demand for drug screening, the heart chip platform has evolved from a single-flux platform to a multi-flux platform using a single perfusion system, but it is not possible to test different experimental conditions at the same time. Therefore, by introducing a tree-like stepped perfusion channel 4 to form different experimental condition groups, efficient drug screening can be achieved.

[0063] In the past, electrical signal monitoring of heart chips mainly relied on patch clamping, which was invasive, disposable, and time-consuming, limiting its application in drug screening. The design of the micro-gold electrode array can significantly increase the flux, achieve non-invasive monitoring, and real-time monitoring. It can also be used as an electrical pacing medium to promote the synchronized beating of myocardial tissue.

[0064] Heterotypic cell co-culture: With the continuous development of interdisciplinary studies in medicine, the evolution of diseases is mutually causal. The heart chip platform explains the mechanism of disease occurrence and development at the cellular level, and the multi-throughput cell culture chamber (this chip consists of 30 culture chambers6) can inoculate different cells at the same time and be used for cell interaction research.

[0065] A method for manufacturing a multi-flux real-time electrical monitoring heart chip, based on the above multi-flux real-time electrical monitoring heart chip, comprises the following steps:

[0066] S1, respectively making a mold having a perfusion layer 1 structure and a mold having a culture chamber layer 2 structure;

[0067] S2, injecting a mixture of PDMS prepolymer A and crosslinking agent B into the mold and curing it to obtain a perfusion layer 1 and a culture chamber layer 2, respectively, and then cleaning and sterilizing them;

[0068] S3, drilling a hole at a designated position in the culture part of the culture chamber layer 2, bonding the culture chamber layer 2 to the glass substrate-electrode plate 3, and then cleaning and sterilizing;

[0069] S4. After inoculating cells in the culture chamber layer 2, place it in an incubator. After the cells adhere to the wall, bond the perfusion layer 1 and the culture chamber layer 2 to form a complete chip system.

[0070] Further optimizing the scheme, in step S1, the method for manufacturing the mold includes the following steps:

[0071] S101, spin coating SU-8 photoresist on the silicon wafer, pre-baking at 95° for 30 min;

[0072] S102, placing the mask on the silicon wafer and then UV-exposing it, followed by post-baking at 95° for 30 minutes;

[0073] S103, using ethyl lactate for development, hardening the film, temperature 120°, time 30 min.

[0074] The scheme was further optimized. In step S2, PDMS prepolymer A and cross-linking agent B were mixed in a ratio of 10:1. After the mixed solution was injected into the mold, vacuum degassing was performed. After degassing, the mixture was cured in an oven at 80° C. for 1 h to obtain a perfusion layer 1 and a culture chamber layer 2.

[0075] The concentration gradient perfusion layer and the culture chamber were processed by using SU8-3035 negative photoresist and polydimethylsiloxane, and the microchip was manufactured according to standard soft lithography and microfabrication methods.

[0076] Preparation of SU8 mold: spin-coat SU-8 photoresist on silicon wafer, pre-bake at 95°C for 30 min, cover the mask on the silicon wafer with UV exposure, post-bake at 95°C for 30 min, use ethyl lactate for development, and harden the film at 120°C for 30 min; pour a 10:1 mixture of PDMS prepolymer A and crosslinker B on the SU8 mold, vacuum degas, and cure in an oven at 80°C for 1 h. Prepare perfusion layer 1 and culture chamber layer 2, punch holes in culture chamber 6 of culture chamber layer 2, and then clean and sterilize for use.

[0077] The culture chamber layer 2 is bonded to the glass substrate-electrode plate 3 through plasma, and then disinfected and sterilized again. After the cells are inoculated in the culture chamber 6 and placed in an incubator, the perfusion layer 1 is bonded to the culture chamber layer 2 after the cells adhere to the wall to form a complete chip system.

[0078] The injection pump is connected to the inlet of the tree-like stepped perfusion channel 4 with a PTFE tube, a flat needle and a syringe to realize fluid drive on the chip: cell culture fluid is injected at the inlet, and the culture fluid automatically forms a drug concentration gradient through the tree-like stepped perfusion channel 4. Multiple drug concentrations can be detected in one experiment, realizing efficient drug screening function.

[0079] Based on the heart chip of the utility model, in situ electrical stimulation of cardiomyocytes and collection of field potential can be performed.

[0080] Drug screening can be performed based on the concentration screening mechanism of the utility model. As one of the application examples, it can be used to detect changes in myocardial cell protein secretion and structural changes in different LDN57444 drug concentration groups under a high uric acid environment.

[0081] (1) The experimental results show that the chip can effectively screen the concentration range of LDN57444 that inhibits the secretion of the inflammatory factor IL-8.

[0082] (2) The experimental results show that the chip can effectively screen the concentration range of LDN57444 that improves the structure of cardiomyocytes. In the range of 0.5-1μm, there is no significant hypertrophy and shrinkage of cardiomyocytes.

[0083] In the description of the present invention, it should be understood that the terms "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside" and "outside" etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0084] The embodiments described above are only descriptions of the preferred methods of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should fall within the protection scope determined by the claims of the present invention.

Claims

1. A multi-flux real-time electrical monitoring heart chip, characterized in that: include: A perfusion layer (1), a culture chamber layer (2) and a glass substrate-electrode plate (3) fixedly connected in sequence from top to bottom; The perfusion layer (1) is provided with a tree-like stepped perfusion channel (4), the tree-like stepped perfusion channel (4) comprising n-level mixing channel groups arranged in sequence and connected along the liquid flow direction, n being a positive integer, the mixing channel group comprising n+1 liquid inlet ends and n+2 liquid outlet ends; the liquid outlet end of the previous mixing channel group corresponds to and is connected to the liquid inlet end of the next mixing channel group; The liquid outlet ends of the mixing channel group located at the tail end are respectively connected to the liquid inlet ends of the liquid injection part, and the liquid outlet ends of the liquid injection part are connected to the outlet (9); The culture chamber layer (2) is provided with a plurality of culture parts, the plurality of culture parts correspond to the plurality of liquid injection parts one by one, and the liquid inlet end of the culture part is connected to the middle of the liquid injection part; A plurality of electrode parts are arranged on the glass substrate-electrode plate (3), and the plurality of electrode parts correspond to and are connected with the plurality of culture parts one by one.

2. A multi-flux real-time electrical monitoring heart chip according to claim 1, characterized in that: The liquid injection portion comprises a plurality of perfusion chambers (5) connected in sequence, wherein the liquid inlet end of the perfusion chamber (5) located at the front end in the liquid flow direction is connected to the liquid outlet end of the corresponding mixing channel group, and the liquid outlet end of the perfusion chamber (5) located at the rear end in the liquid flow direction is connected to the corresponding outlet (9); A plurality of the perfusion chambers (5) are connected to the corresponding culture parts.

3. A multi-flux real-time electrical monitoring heart chip according to claim 2, characterized in that: The culture part comprises a plurality of culture chambers (6), the plurality of culture chambers (6) correspond one to one with the plurality of perfusion chambers (5), the culture chambers (6) are connected with the perfusion chambers (5), and one end of the electrode part is correspondingly arranged in the culture chamber (6).

4. A multi-flux real-time electrical monitoring heart chip according to claim 3, characterized in that: The electrode portion comprises two internal signal collection targets (7), the internal signal collection targets (7) are electrically connected to edge output signal targets (8), and the internal signal collection targets (7) are fixed to the bottom of the corresponding culture chamber (6).

5. The multi-flux real-time electrical monitoring heart chip according to claim 2, characterized in that: The liquid injection part comprises three perfusion chambers (5), the three perfusion chambers (5) are connected in sequence, the perfusion chamber (5) is an elliptical structure of 3.5mm*6mm, and the height of the perfusion chamber (5) is 150um.

6. The multi-flux real-time electrical monitoring heart chip according to claim 3, characterized in that: The culture part comprises three culture chambers (6), each of which is an elliptical structure of 3.5 mm*6 mm, and the thickness of the culture chamber (6) is 2 mm.

Images