CN222524564U - A multi-flux real-time electrical monitoring heart chip - Google Patents
A multi-flux real-time electrical monitoring heart chip Download PDFInfo
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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 chamber layer and a glass substrate-electrode plate, wherein the perfusion layer, the culture chamber layer and the glass substrate-electrode plate 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-level mixed flow channel groups which are sequentially communicated in the liquid flow direction, liquid outlet ends of the mixed flow channel groups at the tail ends are respectively communicated with liquid inlet ends of liquid injection parts, liquid outlet ends of the liquid injection parts are communicated with outlets, a plurality of culture parts are formed in the culture chamber layer, the culture parts are in one-to-one correspondence with the liquid inlet ends of the liquid injection parts, a plurality of electrode parts are arranged on the glass substrate-electrode plate, and the electrode parts are in one-to-one correspondence with and are connected with the culture parts.
Description
Technical Field
The utility model belongs to the technical field of heart chips, and particularly relates to a multi-flux real-time electrical monitoring heart chip.
Background
Heart disease is a significant burden on the global health care system and is also a leading cause of yearly death. In order to improve our understanding of heart disease, a high quality disease model and an efficient drug screening method are required, and along with the rapid development of microfluidic and soft lithography technologies and the progress of human Induced Pluripotent Stem Cell (iPSC) biology, a brand new "organ on chip" field is generated, aiming at establishing a controllable and lowest functional unit capable of reproducing specific organ level functions in vitro.
The method is combined with a microfluidic technology to realize long-term dynamic culture and observation of cells in a closed environment, and combined with a sensing technology (electrical sensing, pressure sensing) and a biological material to monitor myocardial tissue contraction force change through a polymaleic acid octamethylene (anhydride) citrate (POMaC) and a crack sensor integrated cantilever, realize electric signal monitoring through a gold electrode and a platinum wire electrode and realize myocardial contraction visualization through inverse opal hydrogel and a capillary tube. Although the construction of the heart chip platform has a solid foundation, the contradiction between bionics and flux integration is still to be solved urgently, and the efficient drug screening function cannot be realized on a single platform.
Aiming at the problems of low flux, insufficient bionics and the like of the existing heart chip, we provide a multi-flux real-time electrical monitoring heart chip.
Disclosure of utility model
The utility model aims to provide a multi-flux real-time electrical monitoring heart chip so as to solve the problems.
In order to achieve the above object, the present utility model provides the following solutions:
A multi-flux real-time electrical monitoring heart chip comprises a perfusion layer, a culture chamber layer and a glass substrate-electrode plate which are fixedly connected in sequence from top to bottom;
The tree-shaped step perfusion channel comprises n-level mixed flow channel groups which are sequentially communicated in the liquid flowing direction, wherein n is a positive integer, and the mixed flow channel groups comprise n+1 liquid inlet ends and n+2 liquid outlet ends;
The liquid outlet ends of the mixing runner groups at the tail ends are respectively communicated with the liquid inlet ends of the liquid injection parts, and the liquid outlet ends of the liquid injection parts are communicated with an outlet;
a plurality of culture parts are arranged on the culture chamber layer, 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 part of the liquid injection parts;
The glass substrate-electrode plate is provided with a plurality of electrode parts, and the electrode parts are in one-to-one correspondence with and connected with the culture parts.
Preferably, the liquid injection part comprises a plurality of perfusion cavities which are communicated in sequence, the liquid inlet end of the perfusion cavity positioned at the front end of the liquid flow direction is communicated with the liquid outlet end of the corresponding mixing flow channel group, and the liquid outlet end of the perfusion cavity positioned at the rear end of the liquid flow direction is communicated with the corresponding outlet;
the perfusion cavities are communicated with the corresponding culture parts.
Preferably, the culture part comprises a plurality of culture chambers, the culture chambers are in one-to-one correspondence with the perfusion chambers, the culture chambers are communicated with the perfusion chambers, and one ends of the electrode parts are correspondingly arranged in the culture chambers.
Preferably, the electrode part comprises two internal signal acquisition targets, the internal signal acquisition targets are electrically connected with edge output signal targets, and the internal signal acquisition targets are fixed at the bottoms of the corresponding culture chambers.
Preferably, the liquid injection part comprises three perfusion cavities, the three perfusion cavities are sequentially communicated, the perfusion cavities are of an elliptic structure with the thickness of 3.5mm and 6mm, and the height of the perfusion cavities is 150um.
Preferably, the culture part comprises three culture chambers, the culture chambers are of an elliptic structure with the thickness of 3.5mm by 6mm, and the thickness of the culture chambers is 2mm.
A manufacturing method of a multi-flux real-time electrical monitoring heart chip is based on the multi-flux real-time electrical monitoring heart chip, and comprises the following steps:
s1, respectively manufacturing a mold with the perfusion layer structure and a mold with the culture chamber layer structure;
s2, injecting the mixed solution of the PDMS prepolymer A and the cross-linking agent B into the mold, solidifying the mixed solution to obtain the perfusion layer and the culture chamber layer respectively, and cleaning and sterilizing the perfusion layer and the culture chamber layer;
s3, punching holes at designated positions in the culture part of the culture chamber layer, bonding the culture chamber layer with the glass substrate-electrode plate, and then cleaning and sterilizing;
S4, inoculating cells in the culture chamber layer, placing the cells in an incubator, and bonding the perfusion layer and the culture chamber layer to form a complete chip system after the cells are attached to the wall.
Preferably, in the step S1, the method for manufacturing the mold includes the steps of:
S101, spin-coating SU-8 photoresist on a silicon wafer, and pre-baking at the temperature of 95 ℃ for 30min;
S102, covering a mask on a silicon wafer, then performing UV exposure, and performing post-baking for 30min at the temperature of 95 ℃;
s103, developing by using ethyl lactate, hardening at 120 ℃ for 30min.
Preferably, in the step S2, the PDMS prepolymer a and the cross-linking agent B are mixed according to a ratio of 10:1, and after the mixed solution is injected into a mold, vacuum degassing is performed, and after the degassing, the mixture is cured in an oven at 80 ℃ for 1 hour, so as to obtain the perfusion layer and the culture chamber layer.
Compared with the prior art, the utility model has the following advantages and technical effects:
When the heart chip is used, the injection pump is connected with the liquid inlet of the tree-shaped step perfusion channel through the PTFE tube, the flat needle head and the injector, two liquid inlets of the mixed flow channel group at the forefront end of the liquid inflow direction of the tree-shaped step perfusion channel are arranged, one liquid inlet is used for injecting culture medium with specific concentration of medicines, the other liquid inlet is used for injecting culture medium without medicines, when the culture medium with specific concentration of medicines and the culture medium without medicines are injected simultaneously, a medicine concentration gradient is automatically formed under the action of the tree-shaped step perfusion channel, liquid medicine with different concentrations enters the corresponding liquid injection part, the liquid injection part is injected into the corresponding culture part arranged on the culture chamber layer, myocardial cells are cultured in the culture part, after the culture part is filled with the liquid medicine, the liquid medicine flows out through the outlet, and the electrode parts arranged in the culture parts are used for electrically stimulating the myocardial cells and detecting cell field potential.
Drawings
For a clearer description of an embodiment of the utility model or of the solutions of the prior art, the drawings that are needed in the embodiment will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the utility model, and that other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art:
FIG. 1 is a schematic view of a perfusion layer structure according to the present utility model;
FIG. 2 is a schematic view of the layer structure of the culture chamber of the present utility model;
FIG. 3 is a schematic view of a glass substrate-electrode plate structure according to the present utility model;
FIG. 4 is a schematic diagram showing the relative positions of the signal acquisition targets and the culture chambers in the utility model;
FIG. 5 is a concentration gradient profile of the present utility model;
FIG. 6 is a graph showing the outlet concentration profile of the present utility model;
FIG. 7 is a schematic diagram of the structure of the present utility model;
FIG. 8 is a flow chart of the manufacturing process of the utility model;
FIG. 9 is a schematic view of the present utility model in use;
FIG. 10 is a waveform diagram of in situ myocardial cell field potential acquisition in accordance with the present utility model;
FIG. 11 is a functional characterization of concentration screening-detection of secreted proteins of cardiomyocytes according to the present utility model;
The device comprises a perfusion layer 1, a culture chamber layer 2, a glass substrate-electrode plate 3, a tree-shaped stepped perfusion channel 4, a perfusion cavity 5, a culture chamber 6, an internal acquisition signal target point 7, an edge output signal target point 8 and an outlet.
Detailed Description
The following description of the embodiments of the present utility model will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present utility model, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.
In order that the above-recited objects, features and advantages of the present utility model will become more readily apparent, a more particular description of the utility model will be rendered by reference to the appended drawings and appended detailed description.
Referring to fig. 1 to 11, the utility model discloses a multi-flux real-time electrical monitoring heart chip, which comprises a perfusion layer 1, a culture chamber layer 2 and a glass substrate-electrode plate 3 which are fixedly connected in sequence from top to bottom;
The tree-shaped stepped perfusion channel 4 is arranged on the perfusion layer 1, and the tree-shaped stepped perfusion channel 4 comprises n-level mixed flow channel groups which are sequentially communicated in the liquid flow direction, wherein n is a positive integer, and each mixed flow channel group comprises n+1 liquid inlet ends and n+2 liquid outlet ends;
The liquid outlet ends of the mixing runner groups at the tail ends are respectively communicated with the liquid inlet ends of the liquid injection parts, and the liquid outlet ends of the liquid injection parts are communicated with the outlet 9;
a plurality of culture parts are arranged on the culture chamber layer 2, 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 part of the liquid injection parts;
The glass substrate-electrode plate 3 is provided with a plurality of electrode parts, and the electrode parts are in one-to-one correspondence with and connected with the culture parts.
When the heart-shaped cell culture system is used, an injection pump is connected with a liquid inlet of the tree-shaped step perfusion channel 4 through a PTFE tube, a flat needle head and an injector, two liquid inlet ends of a mixed flow channel group at the forefront end of the liquid inflow direction of the tree-shaped step perfusion channel 4 are arranged, one of the liquid inlet ends is used for injecting a culture medium with specific concentration of drugs, the other liquid inlet end is used for injecting a culture medium without the drugs, when the culture medium with the specific concentration of drugs and the culture medium without the drugs are injected simultaneously, a drug concentration gradient is automatically formed under the action of the tree-shaped step perfusion channel 4, liquid medicine with different concentrations is enabled to enter corresponding liquid injection parts, the liquid injection parts are injected into corresponding culture parts arranged on a culture chamber layer 2, myocardial cells are cultured in the culture parts, after the culture parts are filled with the liquid medicine, the liquid medicine flows out through an outlet 9, and the electrode parts arranged in each culture part are used for carrying out electric stimulation and detection of cell field potential on the myocardial cells.
In a further optimization scheme, the liquid injection part comprises a plurality of perfusion cavities 5 which are communicated in sequence, the liquid inlet end of the perfusion cavity 5 positioned at the front end of the liquid flow direction is communicated with the liquid outlet end of the corresponding mixed flow channel group, and the liquid outlet end of the perfusion cavity 5 positioned at the rear end of the liquid flow direction is communicated with the corresponding outlet 9;
the perfusion chambers 5 are communicated with the corresponding culture parts.
Further optimizing scheme, cultivate the portion and include a plurality of cultivate room 6, a plurality of cultivate room 6 and a plurality of perfusion chamber 5 one-to-one, cultivate room 6 and perfusion chamber 5 intercommunication, the one end of electrode portion corresponds the setting in cultivate room 6.
Further optimizing scheme, electrode part includes two inside collection signal targets 7, and inside collection signal targets 7 electric connection has marginal output signal target 8, and inside collection signal targets 7 are fixed in the bottom of corresponding cultivateing room 6.
Aiming at the problems of low flux, insufficient bionics and the like of the existing heart chip, the utility model constructs a microfluidic real-time dynamic monitoring heart chip platform so as to realize flux and bionics mechanism integrated technology, and applies the integrated technology to heart disease modeling, drug screening, cell interaction and other researches, explores the occurrence and development rules and mechanisms of diseases and searches for disease markers and drug targets, thereby providing a new tool for relevant biomedical research.
The utility model is of a layer-by-layer stacked structure, which is divided into 3 parts, wherein the uppermost layer is a perfusion layer 1, a tree-shaped ladder perfusion channel 4 and a plurality of perfusion cavities 5 are arranged on the perfusion layer 1, the perfusion cavities 5 of a culture chamber are of an elliptic structure with the diameter of 3.5mm and the diameter of 6mm, the perfusion cavities 5 are distributed in an array, the height of the perfusion cavities 5 is preferably 150um, 30 perfusion cavities 5 are preferably distributed in 10 rows and 3 columns, the thickness of the perfusion layer 1 is 2mm, the height of the tree-shaped ladder perfusion channel 4 is 150um, the bottom surface of the perfusion layer 1 is in direct contact with the top surface of the culture chamber layer 2, the perfusion cavities 5 are in butt joint and communication with the tops of the corresponding culture chambers 6, the liquid inlet ends of the mixed flow channel group positioned at the forefront end of the liquid inflow direction of the tree-shaped ladder perfusion channel 4 are of 1mm in diameter, the diameter of the outlet 9 is preferably 1mm, after the culture medium containing a specific concentration of medicine and the medicine-free culture medium are respectively filled in the culture medium from the inlet, and the outlet 9 can obtain linear culture medium containing the concentration gradient of the medicine and different cell culture medium for forming the linear gradient.
Referring to fig. 4-5, by comsolmultiphysisc software simulation of this layer, a stable linear concentration gradient can be formed, the right outlet concentration is in a linear relationship from top to bottom, and the simulated data is plotted to obtain fig. 5, and it can be seen from fig. 5 that the outlet concentration is in a linear relationship.
The middle layer is a culture chamber layer 2, a plurality of culture chambers 6 are arranged on the culture chamber layer 2, the culture chambers 6 are of an elliptic structure with the diameter of 3.5mm and 6mm, the culture chambers 6 are distributed in an array, the utility model preferably comprises 30 culture chambers 6, the culture chambers 6 are arranged in one-to-one correspondence with the perfusion chambers 5, the thickness of the culture chambers 6 is 2mm, the width of the culture chambers 6 is 3.5mm, the length is 6mm, the layer is used for inoculating cells for cell culture, the upper sides of the culture chambers 6 are communicated with the corresponding perfusion chambers 5, and the lower sides of the culture chambers 6 are contacted with the glass substrate-electrode plate 3. The independent perfusion cavity 5 and the culture chamber 6 enable the culture medium of the perfusion layer 1 to be soaked and fully fill the culture chamber 6, so that the impact force of perfusion on the adherent cells is reduced.
The glass plate of the glass substrate-electrode plate 3 has a thickness of 1mm, the top surface (the contact surface with the culture chamber layer 2) is firstly plated with titanium with a thickness of 20nm and then plated with gold with a thickness of 50nm, and a microelectrode array is formed, wherein an internal acquisition signal target 7 is a rectangle of 0.4mm multiplied by 0.4mm, an edge output signal target 8 is a rectangle of 1.37mm multiplied by 1.37mm, and the two are connected through a passage with a width of 200 um. The bottom of each culture chamber 6 is provided with two internal signal acquisition targets 7.
The microelectrode array can perform electric stimulation and field potential acquisition in real time in the cell culture process, namely a signal generator is connected with an output signal target point, a stimulation signal is transmitted to an internal acquisition signal target point 7 through the microelectrode array to act on cells to control the pulsation frequency and pulsation amplitude of the cells, otherwise, the microelectrode array can transmit the field potential generated by the pulsation of the cells to an edge output signal target point 8, and the edge output signal target point 8 is connected with a signal amplifier to convert the field potential of the cells into a visual oscillogram.
Compared with the current heart chip, the platform has more multifunction, can realize the high-efficiency drug screening function on a single platform, can realize the co-culture of abnormal cells in a closed system, and improves the real-time electrophysiological evaluation flux.
Multiple flux as drug screening demands increase, cardiac chip platforms have undergone multiple flux platforms that evolve from single flux to single perfusion systems, but testing under different experimental conditions at the same time has not been achieved. Therefore, by introducing the tree-shaped step perfusion channel 4 to form different test condition groups, the efficient drug screening is achieved.
The electrical signal monitoring of the traditional heart chip mainly depends on patch clamp, and the application of the patch clamp in medicine screening is limited due to the invasiveness, disposability and long time consumption. The design of the micro-gold electrode array can obviously improve flux, realize noninvasive monitoring and real-time monitoring, and simultaneously can be used as an electric pacing medium to promote the synchronous beating of myocardial tissues.
Abnormal cell co-culture, that is, along with the continuous development of interdisciplinary subjects in medicine, the evolution of diseases is causal. The heart chip platform expounds the disease occurrence and development mechanism from the cell level, and the multi-flux cell culture chamber (the chip consists of 30 culture chambers 6) can be used for simultaneously inoculating different cells and researching cell interaction.
A manufacturing method of a multi-flux real-time electrical monitoring heart chip is based on the multi-flux real-time electrical monitoring heart chip, and comprises the following steps:
s1, respectively manufacturing a mould with a perfusion layer 1 structure and a culture chamber layer 2 structure;
S2, injecting the mixed solution of the PDMS prepolymer A and the cross-linking agent B into a mould, solidifying to obtain a perfusion layer 1 and a culture chamber layer 2 respectively, and cleaning and sterilizing;
s3, punching holes at designated positions in the culture part of the culture chamber layer 2, bonding the culture chamber layer 2 with the glass substrate-electrode plate 3, and cleaning and sterilizing;
s4, after the cells are received in the culture chamber layer 2, placing the cells in an incubator, and after the cells are attached to the wall, bonding the perfusion layer 1 and the culture chamber layer 2 to form a complete chip system.
In a further optimized scheme, in step S1, the manufacturing method of the mold includes the following steps:
S101, spin-coating SU-8 photoresist on a silicon wafer, and pre-baking at the temperature of 95 ℃ for 30min;
S102, covering a mask on a silicon wafer, then performing UV exposure, and performing post-baking for 30min at the temperature of 95 ℃;
s103, developing by using ethyl lactate, hardening at 120 ℃ for 30min.
In a further optimized scheme, in the step S2, PDMS prepolymer A and cross-linking agent B are mixed according to the proportion of 10:1, the mixed solution is injected into a mould, vacuum degassing is carried out, and the mixture is solidified for 1h in an oven at 80 ℃ after the degassing, so that a perfusion layer 1 and a culture chamber layer 2 are obtained.
The chip processing of the concentration gradient perfusion layer and the culture chamber adopts SU8-3035 negative photoresist and polydimethylsiloxane, and microchip manufacturing is carried out according to standard soft photoetching and micromachining methods.
The preparation method of the SU8 mold comprises the steps of spin coating SU-8 photoresist on a silicon wafer, pre-baking for 95 ℃ for 30min, covering a mask on the silicon wafer, UV exposure, post-baking for 95 ℃ for 30min, developing by using ethyl lactate, hardening for 120 ℃ for 30min, pouring a 10:1 mixture of PDMS prepolymer A and a cross-linking agent B on the SU8 mold, and curing for 1h in an oven at 80 ℃ after vacuum degassing. The perfusion layer 1 and the culture chamber layer 2 are prepared, and the culture chamber 6 of the culture chamber layer 2 is perforated and then cleaned and sterilized for standby.
The culture chamber layer 2 is bonded with the glass substrate-electrode plate 3 through Plasma, and is disinfected and sterilized again, cells are received in the culture chamber 6, then the cells are placed in an incubator, and after the cells are attached to the wall, the perfusion layer 1 is bonded with the culture chamber layer 2 to form a complete chip system.
The PTFE tube, the flat needle head and the injector are connected with the injection pump and the inlet of the tree-shaped step perfusion channel 4 to realize the fluid drive on the chip, wherein the cell culture solution is injected into the inlet, and the culture solution automatically forms a medicine concentration gradient through the tree-shaped step perfusion channel 4, so that the detection of various medicine concentrations can be realized by one experiment, and the high-efficiency medicine screening function is realized.
The heart chip can be used for carrying out myocardial cell in-situ electric stimulation and field potential acquisition.
The concentration screening mechanism can be used for drug screening, and can be used for detecting myocardial cell protein secretion change and structural change of different LDN57444 drug concentration groups under uric acid environment as one application example.
(1) The experimental result shows that the chip can effectively screen the LDN57444 concentration range for inhibiting the secretion of inflammatory factor IL-8.
(2) The experimental result shows that the chip can effectively screen the LDN57444 concentration range for improving the myocardial cell structure, and the myocardial cells have no obvious hypertrophy and shrinkage within the range of 0.5-1 mu m.
In the description of the present utility model, it should be understood that the terms "longitudinal," "transverse," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like indicate or are based on the orientation or positional relationship shown in the drawings, merely to facilitate description of the present utility model, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the present utility model.
The above embodiments are only illustrative of the preferred embodiments of the present utility model and are not intended to limit the scope of the present utility model, and various modifications and improvements made by those skilled in the art to the technical solutions of the present utility model should fall within the protection scope defined by the claims of the present utility model without departing from the design spirit of the present utility model.
Claims (6)
1. The multi-flux real-time electrical monitoring heart chip is characterized by comprising a perfusion layer (1), a culture chamber layer (2) and a glass substrate-electrode plate (3) which are fixedly connected in sequence from top to bottom;
The tree-shaped stepped perfusion channel (4) is formed in the perfusion layer (1), the tree-shaped stepped perfusion channel (4) comprises n-level mixed flow channel groups which are sequentially communicated in the liquid flow direction, n is a positive integer, and the mixed flow channel groups comprise n+1 liquid inlet ends and n+2 liquid outlet ends;
The liquid outlet ends of the mixing runner groups at the tail ends are respectively communicated with the liquid inlet ends of the liquid injection parts, and the liquid outlet ends of the liquid injection parts are communicated with an outlet (9);
A plurality of culture parts are arranged on the culture chamber layer (2), 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 part of the liquid injection parts;
The glass substrate-electrode plate (3) is provided with a plurality of electrode parts, and the electrode parts are in one-to-one correspondence with and connected with the culture parts.
2. The multi-flux real-time electrical monitoring heart chip of claim 1, wherein the liquid injection part comprises a plurality of perfusion cavities (5) which are communicated in sequence, the liquid inlet ends of the perfusion cavities (5) positioned at the front end of the liquid flow direction are communicated with the liquid outlet ends of the corresponding mixing flow channel groups, and the liquid outlet ends of the perfusion cavities (5) positioned at the rear end of the liquid flow direction are communicated with the corresponding outlets (9);
the perfusion cavities (5) are communicated with the corresponding culture parts.
3. The multi-flux real-time electrical monitoring heart chip of claim 2, wherein the culture part comprises a plurality of culture chambers (6), the culture chambers (6) are in one-to-one correspondence with the perfusion chambers (5), the culture chambers (6) are communicated with the perfusion chambers (5), and one ends of the electrode parts are correspondingly arranged in the culture chambers (6).
4. A multi-flux real-time electrical monitoring heart chip according to claim 3, wherein the electrode part comprises two internal collection signal targets (7), the internal collection signal targets (7) are electrically connected with edge output signal targets (8), and the internal collection signal targets (7) are fixed at the bottoms of the corresponding culture chambers (6).
5. The multi-flux real-time electrical monitoring heart chip of claim 2, wherein the liquid injection part comprises three perfusion cavities (5), the three perfusion cavities (5) are sequentially communicated, the perfusion cavities (5) are of an elliptic structure with the size of 3.5mm by 6mm, and the height of the perfusion cavities (5) is 150um.
6. A multi-flux real-time electrical monitoring heart chip according to claim 3, wherein the culture part comprises three culture chambers (6), wherein the culture chambers (6) are of an elliptical structure of 3.5mm by 6mm, and the thickness of the culture chambers (6) is 2mm.
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