System and method for detecting spatial trans-epithelial impedance

Abstract

The invention discloses a detection system and method for spatial trans-epithelial impedance, and relates to the field of biomedical engineering.The detection system for spatial trans-epithelial impedance comprises a cell culture pool and a detector; the cell culture pool comprises a culture device, an upper electrode, a lower electrode array and a thin film, the thin film divides the culture device into an upper culture device and a lower culture device, and the upper culture device is used for growing epithelial cells above the thin film; one end of the upper electrode is inserted into the upper culture solution of the upper incubator, and the other end is connected with the detector; the lower electrode array is located at the bottom of the lower incubator and connected with the detector through a metal lead, and the height of the lower-layer culture solution of the lower incubator is at least larger than or equal to that of the film; a circuit channel is formed between each lower electrode and the upper electrode in the lower electrode array; according to the system, the space solution of the trans-epithelial impedance can be realized by combining the detector with the equivalent circuit of the cell culture pool, and the difference between the trans-epithelial impedance of different spaces can be reflected.

Description

Technical Field

[0001] The present invention relates to the field of biomedical engineering, and in particular to a detection system and method of spatial transepithelial impedance. Background Art

[0002] Cell barriers play a key role in living organisms, such as maintaining the integrity of tissue structures, regulating the transport of substances and the communication between cells. Understanding the properties and functions of cell barriers is of great significance for understanding the mechanisms of disease onset, drug delivery and biomaterials. Transepithelial electrical resistance (TEER) is a label-free measurement method that is commonly used to assess the integrity and tightness of tissue barriers. Common TEER measurement methods are achieved through organ-on-chip structures, for example, by inserting metal electrodes into designated locations.

[0003] However, this method requires a relatively long culture cycle and device design. At the same time, due to the structure and materials of the organ chip, its electrodes are relatively simple, and the spatial distribution of TEER cannot be obtained. Although a monolayer of cells can be obtained in the channel, it is still difficult to obtain barrier information at a specific location, because the barrier is generally evaluated based on the impedance information of TEER in the time dimension, which makes it lack information on the spatial dimension. At the same time, since the traditional TEER monitoring technology is the average impedance value of a large area of ​​cell area, the accuracy of the result will also be affected by the area with the highest conductivity in the entire cell area. The TEER value reflects the different states of the cell layer (drug effect), and its accuracy will affect the results of the cell layer state it reflects.

[0004] Therefore, how to perform spatial detection of transepithelial impedance has become a technical problem that needs to be solved urgently by those skilled in the art. Summary of the invention

[0005] The object of the present invention is to provide a spatial transepithelial impedance detection system and method, so as to perform spatial detection of transepithelial impedance.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention first provides a spatial transepithelial impedance detection system, comprising:

[0008] A spatial trans-epithelial impedance detection system, characterized in that it comprises: a cell culture pool, a detector; the cell culture pool comprises: a culture vessel, an upper electrode, a lower electrode array, and a thin film, the thin film separates the culture vessel into an upper culture vessel and a lower culture vessel, the upper culture vessel is used to grow epithelial cells above the thin film; one end of the upper electrode is inserted into the upper culture fluid in the upper culture vessel, and the other end is connected to the detector; the lower electrode array is located at the bottom of the lower culture vessel and is connected to the detector through a metal lead, the height of the lower culture fluid in the lower culture vessel is at least greater than or equal to the height of the thin film; each lower electrode in the lower electrode array and the upper electrode respectively form a circuit channel;

[0009] The detector is used to obtain the background impedance of each of the circuit channels when the epithelial cells are not grown in the cell culture tank; obtain the first transverse equivalent impedance of the transverse equivalent circuit of adjacent lower electrodes of the lower electrode array in the lower culture fluid; and, when the epithelial cells are grown in the cell culture tank, obtain the total impedance of each of the circuit channels; determine the second transverse equivalent impedance of the circuit channel by combining the first transverse equivalent impedance and the position of the lower electrode corresponding to the circuit channel in the lower electrode array; and determine the spatial transepithelial impedance corresponding to each of the circuit channels based on the total impedance of each of the circuit channels, the background impedance and the second transverse equivalent impedance.

[0010] In an optional embodiment of the present application, when the epithelial cells are not grown in the cell culture pool, obtaining the background impedance of each of the circuit channels includes:

[0011] When the epithelial cells are not grown in the cell culture tank, obtaining an EIS impedance spectrum of each circuit channel of the cell culture tank;

[0012] Determine, according to the EIS impedance spectrum, the electrical double layer impedance of the electrode surface of the upper electrode and the lower electrode of each of the circuit channels, the impedance of the upper culture solution, the impedance of the film, and the longitudinal equivalent impedance of the lower culture solution corresponding to each of the circuit channels;

[0013] The background impedance of each circuit channel is determined according to the electrical double layer impedance of the electrode surface, the impedance of the upper culture solution, the impedance of the film and the longitudinal equivalent impedance.

[0014] In an optional implementation manner of the present application, the background impedance of each of the circuit channels is expressed by the following formula:

[0015] Z F =2*Z CPE +R up +RB +R M ;

[0016] Among them, Z F represents the background impedance; Z CPE represents the electrical double layer impedance of the electrode surface; R up Represents the impedance of the upper culture solution; R M represents the impedance of the film; R B Denotes the longitudinal equivalent impedance.

[0017] In an optional embodiment of the present application, obtaining a first lateral equivalent impedance of a lateral equivalent circuit of adjacent lower electrodes of the lower electrode array in the lower culture solution includes:

[0018] The first lateral equivalent impedance is obtained by scanning the EIS impedance spectrum through adjacent circuit channels.

[0019] In an optional implementation manner of the present application, the determining the second lateral equivalent impedance of the circuit channel in combination with the first lateral equivalent impedance and the position of the lower electrode corresponding to the circuit channel in the lower electrode array includes:

[0020] Determining the position of each of the lower electrodes in the lower electrode array;

[0021] Obtaining impedance coefficients corresponding to any two lower electrodes according to positions of the lower electrodes in the lower electrode array;

[0022] A second transverse equivalent impedance of the circuit path is determined based on the impedance coefficient and the first transverse equivalent impedance.

[0023] In an optional implementation manner of the present application, determining the spatial transepithelial impedance corresponding to each of the circuit channels according to the total impedance of each of the circuit channels, the background impedance, and the second transverse equivalent impedance includes:

[0024] Constructing a correlation between the transepithelial impedance and the total impedance of each circuit channel, the background impedance, and the second transverse equivalent impedance;

[0025] Based on the association relationship, the spatial transepithelial impedance corresponding to each of the circuit channels is obtained.

[0026] In an optional implementation manner of the present application, the association relationship can be expressed by the following formula:

[0027]

[0028] C=B -1 ×A;

[0029] Among them, Z UX Represents the total impedance of the Xth circuit channel; Z F represents the background impedance; Z X represents the spatial transepithelial impedance corresponding to the Xth circuit channel; λ N～M Indicates the intermediate coefficient; R N～M represents the second lateral equivalent impedance corresponding to the circuit channel composed of the lower electrode in the Nth row and the Mth column in the lower electrode array and the upper electrode; N and M are positive integers less than X.

[0030] In an optional embodiment of the present application, R N～M Obtained by the following formula:

[0031] R N～M =K N～M ×R0;

[0032] Among them, K N～M Represents the impedance coefficient corresponding to the Nth lower electrode to the Mth lower electrode; R0 represents the first lateral equivalent impedance. Compared with the prior art, the present invention provides a detection system for spatial transepithelial impedance, which designs a lower electrode array in the lower culture vessel of a cell culture pool, and combines the background impedance of the circuit channel formed between each lower electrode in the lower electrode array and the upper electrode, the total impedance of each circuit channel, and the equivalent circuit of the lower culture fluid in the lower culture vessel to determine the spatial transepithelial impedance corresponding to each circuit channel, thereby realizing the spatial solution of the transepithelial impedance, which is beneficial to reflecting the difference between transepithelial impedances in different spaces.

[0033] The present invention also provides a method for detecting spatial transepithelial impedance, which is applied to the above-mentioned spatial transepithelial impedance detection system, comprising:

[0034] A method for detecting spatial transepithelial impedance, applied to the above-mentioned spatial transepithelial impedance detection system, comprising:

[0035] In the case where no epithelial cells are grown in the cell culture tank, the background impedance of each circuit channel of the cell culture tank is obtained; and the first lateral equivalent impedance of the lateral equivalent circuit of the adjacent lower electrodes of the lower electrode array of the cell culture tank in the lower culture solution is obtained;

[0036] When the epithelial cells are grown in the cell culture tank, obtaining the total impedance of each of the circuit channels;

[0037] Determine, in combination with the first lateral equivalent impedance and the circuit structure of the lateral equivalent circuit, a second lateral equivalent impedance of any two lower electrodes in the lower electrode array in the lateral equivalent circuit;

[0038] The spatial transepithelial impedance corresponding to each of the circuit channels is determined according to the total impedance of each of the circuit channels, the background impedance and the second transverse equivalent impedance.

[0039] Compared with the prior art, the beneficial effects of the spatial transepithelial impedance detection method provided by the present invention are the same as the beneficial effects of the spatial transepithelial impedance detection system described in the above technical solution, which will not be described in detail here.

[0040] The present invention also provides an electronic device, comprising:

[0041] processor;

[0042] a memory for storing instructions executable by the processor;

[0043] The processor is used to execute the above-mentioned spatial transepithelial impedance detection method by running the instructions in the memory.

[0044] Compared with the prior art, the beneficial effects of the electronic device provided by the present invention are the same as the beneficial effects of the spatial transepithelial impedance detection system described in the above technical solution, and will not be described in detail here.

[0045] The present invention also provides a computer storage medium, in which instructions are stored. When the instructions are executed, the above-mentioned spatial transepithelial impedance detection method is implemented.

[0046] Compared with the prior art, the beneficial effects of the computer storage medium provided by the present invention are the same as the beneficial effects of the spatial transepithelial impedance detection system described in the above technical solution, and will not be elaborated here. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0048] Figure 1 A structural diagram of a spatial transepithelial impedance detection system provided in an embodiment of the present application;

[0049] Figure 2 A schematic diagram of the structure of the lower electrode array provided in an embodiment of the present application;

[0050] Figure 3 A schematic diagram of an equivalent circuit of a cell culture tank provided in an embodiment of the present application;

[0051] Figure 4 An equivalent circuit structure diagram provided for an embodiment of the present application;

[0052] Figure 5A structural diagram of a lateral equivalent circuit of a lower culture solution provided in an embodiment of the present application;

[0053] Figure 6 A flow chart of a method for detecting spatial transepithelial impedance provided in an embodiment of the present application;

[0054] Figure 7 A schematic diagram of the structure of an electronic device provided in an embodiment of the present application. DETAILED DESCRIPTION

[0055] In order to clearly describe the technical solutions of the embodiments of the present invention, in the embodiments of the present invention, words such as "first" and "second" are used to distinguish the same items or similar items with basically the same functions and effects. For example, the first threshold and the second threshold are only used to distinguish different thresholds, and their order is not limited. Those skilled in the art can understand that words such as "first" and "second" do not limit the quantity and execution order, and words such as "first" and "second" do not necessarily limit them to be different.

[0056] It should be noted that, in the present invention, words such as "exemplary" or "for example" are used to indicate examples, illustrations or descriptions. Any embodiment or design described as "exemplary" or "for example" in the present invention should not be interpreted as being more preferred or more advantageous than other embodiments or designs. Specifically, the use of words such as "exemplary" or "for example" is intended to present related concepts in a specific way.

[0057] In the present invention, "at least one" means one or more, and "plurality" means two or more. "And / or" describes the association relationship of associated objects, indicating that three relationships may exist. For example, A and / or B can mean: A exists alone, A and B exist at the same time, and B exists alone, where A and B can be singular or plural. The character " / " generally indicates that the associated objects before and after are in an "or" relationship. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items. For example, at least one of a, b or c can mean: a, b, c, the combination of a and b, the combination of a and c, the combination of b and c, or the combination of a, b and c, where a, b, c can be single or multiple.

[0058] The present application provides a system and method for detecting spatial transepithelial impedance, which are described in detail one by one in the following embodiments.

[0059] The present application embodiment first provides a spatial transepithelial impedance detection system, please refer to Figure 1 , Figure 1 This is a structural diagram of the spatial transepithelial impedance detection system provided in an embodiment of the present application.

[0060] like Figure 1 As shown, the spatial transepithelial impedance detection system comprises: a cell culture pool 101 and a detector 102;

[0061] The cell culture pool 101 includes: a culture vessel 1, an upper electrode 2, a lower electrode array 3, a film 4, epithelial cells 5, an upper culture solution 6, and a lower culture solution 7.

[0062] The film 4 separates the culture vessel 1 into an upper culture vessel and a lower culture vessel, and the upper culture vessel is used to grow epithelial cells 5 above the film 4; one end of the upper electrode is inserted into the upper culture fluid 6 in the upper culture vessel, and the other end is connected to the detector 102; the lower electrode array 3 is located at the bottom of the lower culture vessel and is connected to the detector 102 through a metal lead; the height of the lower culture fluid 7 is at least greater than or equal to the height of the film 4. Each lower electrode in the lower electrode array 3 forms a circuit channel with the upper electrode 2.

[0063] For the lower electrode array 3, please refer to Figure 2 , Figure 2 A schematic diagram of the structure of the lower electrode array provided in an embodiment of the present application.

[0064] like Figure 2 As shown, the lower electrode array 3 is arranged in a 4×4 matrix, with a total of 16 electrodes, the size of a square with a side length of 0.9 mm and a spacing of 0.1 m. In the actual application process, the lower electrode array can be obtained by wet etching technology. First, 20 nm chromium is evaporated and deposited on the glass substrate as an adhesion layer, and then 200 nm gold is deposited. The wet etching protection layer is realized by spin coating photoresist, exposure, and development. Then, potassium iodide (KI), iodine (I2) and water (DI) are used to prepare a gold etching solution in a ratio of 4g:1g:40g, and the gold is etched for 45s. Then, cerium ammonium nitrate, acetic acid and water (DI) are used to prepare a chromium etching solution in a ratio of 20g:10g:70g, and the etching is carried out for 30s. After the patterning process, the sample is cleaned with acetone to eliminate any residual photoresist.

[0065] It can be understood that the above description of the lower electrode array 3 is only for the purpose of facilitating the understanding of the specific structure and preparation process of the lower electrode array provided in the embodiment of the present application. In actual application, the lower electrode array can select a suitable size according to actual conditions. For example, the lower electrode array 3 can also be an electrode array of sizes such as 5×5 or 3×4, and the present application does not impose any restrictions on this.

[0066] As for the film 4, the film 4 can be made of materials such as PET (Polyethylene Terephthalate) film, and this application does not make any limitation.

[0067] For epithelial cells 5, A549 cells preserved in Dulbecco's modified Eagle medium (DMEM, Biological Industries) can be used, supplemented with 10% heat-inactivated fetal bovine serum (FBS, Biological Industries), 1% glutamine (Gibco) and 1% penicillin-streptomycin-amphotericin B (PSA, Biological Industries) solution, and cultured in a constant temperature and humidity incubator at 37°C and 5% CO2, and then implanted into cell culture pool 101 for monolayer cell culture.

[0068] After the cell culture pool 101 is constructed, the spatial transepithelial impedance corresponding to each circuit channel can be determined by the detector 102 to achieve the detection of the spatial transepithelial impedance.

[0069] Specifically, the detector 102 is used to obtain the background impedance of each of the circuit channels when the epithelial cells are not grown in the cell culture pool; obtain the first transverse equivalent impedance of the transverse equivalent circuit of adjacent lower electrodes of the lower electrode array in the lower culture fluid; and, when the epithelial cells are grown in the cell culture pool, obtain the total impedance of each of the circuit channels; determine the second transverse equivalent impedance of the circuit channel by combining the first transverse equivalent impedance and the position of the lower electrode corresponding to the circuit channel in the lower electrode array; and determine the spatial transepithelial impedance corresponding to each of the circuit channels based on the total impedance of each of the circuit channels, the background impedance and the second transverse equivalent impedance.

[0070] In order to facilitate understanding of the function of the detector 102, the following Figure 3 The detection process of spatial transepithelial impedance by the detector 102 is described.

[0071] Please refer to Figure 3 , Figure 3 This is a schematic diagram of an equivalent circuit of a cell culture tank provided in an embodiment of the present application.

[0072] like Figure 3 As shown, the equivalent circuit of the cell culture pool includes: an epithelial cell equivalent circuit 301, a thin film equivalent circuit 302, a longitudinal equivalent circuit 303 of the lower culture fluid 7, a lateral equivalent circuit 304 of the lower culture fluid 7, an equivalent circuit 305 of the upper culture fluid 6; and an equivalent circuit 306 of the upper electrode and lower electrode arrays.

[0073] Among them, the equivalent circuit 306 of the upper electrode and the lower electrode array can be regarded as the double-layer capacitance of the two electrode surfaces. Due to the double-layer effect, there is a double-layer capacitance on the electrode surface, and its impedance changes with frequency and is expressed as a mixed element of resistance and capacitance. Usually, it is approximated as a constant phase element (CPE), and its impedance can be expressed as the double-layer impedance Z of the electrode surface CPE .

[0074] For the equivalent circuit 305 of the upper culture solution 6, its impedance can be expressed as the impedance R of the upper culture solution up .

[0075] For the film equivalent circuit 302, its impedance can be expressed as the film impedance R M In practical applications, the equivalent impedance R of a film with a space of 0.5 μm and a thickness of 5 μm is M It is about 20Ω, which is much lower than the equivalent impedance of other parts.

[0076] The impedance of the equivalent circuit of the lower culture fluid 7 is analyzed differently in the case where epithelial cells are grown and in the case where epithelial cells are not grown. In the case where epithelial cells are not grown, the equivalent circuit of the lower culture fluid 7 can be regarded as the longitudinal equivalent impedance R of the longitudinal equivalent circuit 303. B .

[0077] In the case of no epithelial cells growing, the first equivalent impedance Z is obtained CPE , the second equivalent impedance R up , the third equivalent impedance R M And the longitudinal equivalent impedance R B , the background impedance of each circuit channel can be determined.

[0078] Specifically, the background impedance of each of the circuit channels can be expressed by the following formula (1):

[0079] Z F =2*Z CPE +R up +R B +R M (1);

[0080] Among them, Z F represents the background impedance; Z CPE represents the electrical double layer impedance of the electrode surface; R up Represents the impedance of the upper culture solution; R M represents the impedance of the film; R B Denotes the longitudinal equivalent impedance.

[0081] In practical applications, the electrical double layer impedance Z CPE, the impedance of the upper culture medium R up , the impedance of the film R M And the longitudinal equivalent impedance R B , can be calculated by scanning the EIS impedance spectrum of each electrode. The EIS impedance spectrum can be obtained using an electrochemical workstation or other resistance or voltage detection devices or equipment. Specifically, the constant current scanning mode can be selected, with a current of 10uA and a frequency range of 10 3 Hz to 10 5 Hz, all analyzed impedance data were impedance data at 10 kHz, to avoid the influence of the electrode surface electric double layer on impedance at low frequency, and to perform multi-channel continuous long-term scanning.

[0082] In the case of growing epithelial cells, the equivalent circuit of the lower culture fluid 7 includes: a longitudinal equivalent circuit 303 and a transverse equivalent circuit 304. The impedance of the transverse equivalent circuit 304 of different lower electrodes in the lower electrode array is also different. The impedance of a resistor in the transverse equivalent circuit 304 of the lower culture fluid can be expressed as R A (i.e., the first lateral equivalent impedance of the lateral equivalent circuit of the adjacent lower electrode in the lower culture solution), the longitudinal equivalent circuit can be expressed as R B .

[0083] Through theoretical calculation, it is expected that R A and R B The relationship between R A / R B >10, assuming that the height of the lower electrode array is 0.2 mm and the distance between adjacent lower electrodes is 1 mm, then by estimation and measurement, it is found that R B About 400Ω, R A It is about 4000Ω, which is an order of magnitude different from most organ chip electrical models in the prior art where the lateral impedance and the longitudinal impedance are of the same order of magnitude. The prior art studies on the impedance of the lower culture fluid are usually based on a single electrode. However, when there are multiple lower electrodes, for a circuit channel consisting of an upper electrode and a lower electrode in a lower electrode array, its total impedance will be affected by the epithelial cells in the area directly above the lower electrode (equivalent to the area corresponding to the area of ​​a lower electrode).

[0084] Take two circuit channels as an example, please refer to Figure 4 , Figure 4 An equivalent circuit structure diagram provided for an embodiment of the present application.

[0085] like Figure 4 As shown, Figure 4 In the figure, U represents the upper electrode 2, X represents the first lower electrode in the lower electrode array 3, Y represents the second lower electrode in the lower electrode array 3, and R Brepresents the longitudinal equivalent circuit of the lower culture medium, R A represents the lateral equivalent circuit of the lower culture medium, Z X represents the transepithelial impedance of the epithelial cells corresponding to the first lower electrode; Z Y represents the transepithelial impedance of the epithelial cells corresponding to the second lower electrode.

[0086] Usually, if the cell layer grows completely uniformly, Z X and Z Y are equal, however, when cells grow unevenly, Z X and Z Y There will also be differences. Therefore, it is necessary to calculate Z X and Z Y When considering the lateral equivalent circuit impedance R A .

[0087] For further information, please refer to Figure 5 , Figure 5 A structural diagram of a lateral equivalent circuit of a lower culture solution provided in an embodiment of the present application.

[0088] Figure 5 The lateral equivalent circuit of the lower culture solution is equivalent to the resistor connected in series between the lower electrodes, and the impedance of the resistor can be expressed as R0. In the embodiment of the present application, the impedance of the resistor is called the first lateral equivalent impedance. In the actual application, R0 can also be determined by EIS impedance spectrum.

[0089] Furthermore, the second transverse equivalent impedance of the circuit channel can be determined by the impedance coefficient corresponding to the circuit channel and R0.

[0090] Specifically, the impedance coefficient corresponding to the circuit channel can be determined by the position of each lower electrode in the lower electrode array:

[0091] Here, assuming that the lower electrode array is 4×4, the lateral equivalent impedance of the circuit channel formed by the lower electrode and the upper electrode located in the first row and first column of the lower electrode array can be expressed as R 1～1 =K 1～1 ×R0; the lateral equivalent impedance of the circuit channel formed by the lower electrode and the upper electrode located in the first row and second column can be expressed as R 1～2 =K 1～2 × R0, ..., and so on, until the second lateral equivalent impedance of all circuit channels is obtained.

[0092] Specifically, the coefficient factor K N～M It can be expressed by the following Table 1:

[0093] Table 1:

[0094] 1 2 3 4 1 0 0.588 0.460 0.382 2 0.588 0.536 0.467 0.407 3 0.460 0.467 0.436 0.391 4 0.382 0.407 0.391 0.350

[0095] It can be understood that in the circuit channel formed by the lower electrode in the first row and the first column of the lower electrode array and the upper electrode, the total impedance Z of the circuit channel is measured. U1 When the lower layer of culture fluid is at 100 μm, the equivalent impedance is equivalent to the spatial transepithelial resistance of 16 circuit channels in parallel (corresponding to the spatial transepithelial impedance Z X ) and the lateral equivalent circuit (the corresponding impedance is the second lateral equivalent impedance R N-M ), then the series background impedance Z F , the final total impedance Z U1 It is expressed by the following formula (2):

[0096] Z U1 =Z F +(Z1 -1 +(Z2+R 1-2 ) -1 +(Z3+R 1-3 ) -1 +(Z4+R 1-4 ) -1 +…+(Z X +R 1-X ) -1 +…+(Z 16 +R 1-16 ) -1 ) -1 (2);

[0097] Among them, Z1 to Z 16 Represents the spatial transepithelial impedance of individual circuit pathways.

[0098] After transforming the above formula (2), the following formula (3) can be obtained:

[0099]

[0100] The total impedance Z measured in each circuit channel is obtained UX and background impedance Z F After the expression is obtained, the total impedance of each circuit channel and the background impedance, as well as the correlation between the second lateral equivalent resistance and the transepithelial impedance can be summarized and constructed.

[0101] Right now,

[0102] C=B -1 ×A;

[0103] Among them, Z UXRepresents the total impedance of the Xth circuit channel; Z F represents the background impedance; Z X represents the spatial transepithelial impedance corresponding to the Xth circuit channel; λ N～M Indicates the intermediate coefficient; R N～M represents the second lateral equivalent impedance corresponding to the circuit channel composed of the lower electrode in the Nth row and the Mth column in the lower electrode array and the upper electrode; N and M are positive integers less than X.

[0104] During the application of the spatial transepithelial impedance detection system, A549 cells were monitored for a long time, and the transepithelial impedance of each space measured for 72 hours was solved. The cell area was stained with calcein, and the staining results were solved by relevant algorithms to obtain the cell coverage of each area. After comparison, it was found that the cell coverage of transepithelial impedance of each circuit channel was highly consistent. The test results show that within the first 24 hours, the transepithelial impedance of different circuit channels did not change significantly. However, after 24 to 96 hours, the transepithelial impedance of different circuit channels began to differ significantly. This is because cell growth is random, so the cell coverage corresponding to different circuit channels varies greatly, especially about 72 hours after inoculation. This highlights the advantages of the spatial transepithelial impedance detection system provided by the present application, which shows that the system can reveal the spatial distribution of cell growth.

[0105] In summary, the spatial transepithelial impedance detection system provided in the embodiment of the present application designs a lower electrode array in the lower culture vessel of the cell culture pool, and combines the background impedance of the circuit channel formed between each lower electrode in the lower electrode array and the upper electrode, the total impedance of each circuit channel, and the equivalent circuit of the lower culture fluid in the lower culture vessel to determine the spatial transepithelial impedance corresponding to each circuit channel, thereby realizing the spatial solution of the transepithelial impedance, which is conducive to reflecting the differences between transepithelial impedances in different spaces.

[0106] Corresponding to the above system embodiment, the present application embodiment also provides a method for detecting spatial transepithelial impedance, which is applied to the spatial transepithelial impedance detection system in the above system embodiment, please refer to Figure 6 , Figure 6 A flow chart of a method for detecting spatial transepithelial impedance provided in an embodiment of the present application.

[0107] like Figure 6 As shown, the method for detecting spatial transepithelial impedance comprises:

[0108] S601, obtaining background impedances of various circuit channels of the cell culture pool when no epithelial cells are grown in the cell culture pool; and obtaining first lateral equivalent impedances of lateral equivalent circuits of adjacent lower electrodes of the lower electrode array of the cell culture pool in the lower culture solution;

[0109] S602, obtaining the total impedance of each of the circuit channels when the epithelial cells are grown in the cell culture pool;

[0110] S603, determining a second transverse equivalent impedance of the circuit channel in combination with the first transverse equivalent impedance and a position of the lower electrode corresponding to the circuit channel in the lower electrode array;

[0111] S604: Determine the spatial transepithelial impedance corresponding to each of the circuit channels according to the total impedance of each of the circuit channels, the background impedance, and the second transverse equivalent impedance.

[0112] In an optional embodiment of the present application, when no epithelial cells are grown in the cell culture pool, obtaining the background impedance of each circuit channel of the cell culture pool comprises:

[0113] When the epithelial cells are not grown in the cell culture tank, obtaining an EIS impedance spectrum of each circuit channel of the cell culture tank;

[0114] Determine, according to the EIS impedance spectrum, the electrical double layer impedance of the electrode surface of the upper electrode and the lower electrode of each of the circuit channels, the impedance of the upper culture solution, the impedance of the film, and the longitudinal equivalent impedance of the lower culture solution corresponding to each of the circuit channels;

[0115] The background impedance of each circuit channel is determined according to the electrical double layer impedance of the electrode surface, the impedance of the upper culture solution, the impedance of the film and the longitudinal equivalent impedance.

[0116] In an optional implementation manner of the present application, the background impedance of each of the circuit channels is expressed by the following formula:

[0117] Z F =2*Z CPE +R up +R B +R M ;

[0118] Among them, Z F represents the background impedance; Z CPE represents the electrical double layer impedance of the electrode surface; R up Represents the impedance of the upper culture solution; R M represents the impedance of the film; RB Denotes the longitudinal equivalent impedance.

[0119] In an optional embodiment of the present application, obtaining a first lateral equivalent impedance of a lateral equivalent circuit of adjacent lower electrodes of the lower electrode array of the cell culture tank in the lower culture solution includes:

[0120] The first lateral equivalent impedance is obtained by scanning the EIS impedance spectrum through adjacent circuit channels.

[0121] In an optional implementation manner of the present application, the determining the second lateral equivalent impedance of the circuit channel in combination with the first lateral equivalent impedance and the position of the lower electrode corresponding to the circuit channel in the lower electrode array includes:

[0122] Determining the position of each of the lower electrodes in the lower electrode array;

[0123] Obtaining impedance coefficients corresponding to any two lower electrodes according to positions of the lower electrodes in the lower electrode array;

[0124] A second transverse equivalent impedance of the circuit path is determined based on the impedance coefficient and the first transverse equivalent impedance.

[0125] In an optional implementation manner of the present application, determining the spatial transepithelial impedance corresponding to each of the circuit channels according to the total impedance of each of the circuit channels, the background impedance, and the second transverse equivalent impedance includes:

[0126] Constructing a correlation between the transepithelial impedance and the total impedance of each circuit channel, the background impedance, and the second transverse equivalent impedance;

[0127] Based on the association relationship, the spatial transepithelial impedance corresponding to each of the circuit channels is obtained.

[0128] In an optional implementation manner of the present application, the association relationship can be expressed by the following formula:

[0129]

[0130] C=B -1 ×A;

[0131] Among them, Z UX Represents the total impedance of the Xth circuit channel; Z F represents the background impedance; Z X represents the spatial transepithelial impedance corresponding to the Xth circuit channel; λ N～M Indicates the intermediate coefficient; R N～Mrepresents the second lateral equivalent impedance corresponding to the circuit channel composed of the lower electrode in the Nth row and the Mth column in the lower electrode array and the upper electrode; N and M are positive integers less than X.

[0132] In an optional embodiment of the present application, R N～M Obtained by the following formula:

[0133] R N～M =K N～M ×R0;

[0134] Among them, K N～M represents the impedance coefficient corresponding to the Nth lower electrode to the Mth lower electrode; R0 represents the first lateral equivalent impedance.

[0135] The spatial transepithelial impedance detection method provided in this embodiment belongs to the same application concept as the spatial transepithelial impedance detection system provided in the above embodiment of this application, and has the beneficial effects of the spatial transepithelial impedance detection method. For technical details not fully described in this embodiment, please refer to the specific processing content of the spatial transepithelial impedance detection method system provided in the above embodiment of this application, which will not be repeated here.

[0136] The present application also provides an electronic device, such as Figure 7 As shown, Figure 7 A schematic diagram of the structure of an electronic device provided in an embodiment of the present application.

[0137] like Figure 7 As shown, the electronic device comprises:

[0138] Processor 210;

[0139] A memory 200 for storing instructions executable by the processor 210;

[0140] The processor 210 is used to execute the spatial transepithelial impedance detection method disclosed in any of the above embodiments by running the instructions in the memory 200.

[0141] The processor 210, the memory 200, the communication interface 220, the input device 230 and the output device 240 are connected to each other via a bus.

[0142] A bus may include a pathway that transfers information between components of a computer system.

[0143] The processor 210 may be a general-purpose processor, such as a general-purpose central processing unit (CPU), a microprocessor, etc., or an application-specific integrated circuit (ASIC), or one or more integrated circuits for controlling the execution of the program of the scheme of the present invention. It may also be a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field programmable gate array (FPGA) or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components.

[0144] The processor 210 may include a main processor, and may also include a baseband chip, a modem, and the like.

[0145] The memory 200 stores a program for executing the technical solution of the present invention, and may also store an operating system and other key services. Specifically, the program may include a program code, and the program code includes a computer operation instruction. More specifically, the memory 200 may include a read-only memory (ROM), other types of static storage devices that can store static information and instructions, a random access memory (RAM), other types of dynamic storage devices that can store information and instructions, a disk storage, a flash, and the like.

[0146] The input device 230 may include a device for receiving data and information input by a user, such as a keyboard, a mouse, a camera, a scanner, a touch screen, etc.

[0147] Output device 240 may include devices that allow information to be output to a user, such as a display screen, printer, speaker, etc.

[0148] The communication interface 220 may include any transceiver or the like to communicate with other devices or communication networks, such as Ethernet, Radio Access Network (RAN), Wireless Local Area Network (WLAN), etc.

[0149] The processor 210 executes the program stored in the memory 200 and calls other devices, and can be used to implement each step of any one of the spatial transepithelial impedance detection methods provided in the above embodiments of the present application.

[0150] In addition to the above methods and devices, embodiments of the present application may also be a computer program product, which includes computer program instructions, which, when executed by a processor, enable the processor to perform the steps in the spatial transepithelial impedance detection method of various embodiments of the present application.

[0151] The computer program product may be written in any combination of one or more programming languages ​​to write program codes for performing the operations of the embodiments of the present application, including object-oriented programming languages, such as Java, C++, etc., and conventional procedural programming languages, such as "C" language or similar programming languages. The program code may be executed entirely on the user computing device, partially on the user device, as an independent software package, partially on the user computing device and partially on a remote computing device, or entirely on a remote computing device or server.

[0152] In addition, the embodiments of the present application may also be a storage medium on which a computer program is stored, and the computer program is executed by a processor to execute the steps in the spatial transepithelial impedance detection method of various embodiments of the present application.

[0153] For the aforementioned method embodiments, for the sake of simplicity, they are all described as a series of action combinations, but those skilled in the art should be aware that the present application is not limited by the order of the actions described, because according to the present application, some steps can be performed in other orders or simultaneously. Secondly, those skilled in the art should also be aware that the embodiments described in the specification are all preferred embodiments, and the actions and modules involved are not necessarily required by the present application.

[0154] It should be noted that each embodiment in this specification is described in a progressive manner, and each embodiment focuses on the differences from other embodiments, and the same or similar parts between the embodiments can be referred to each other. For the device embodiment, since it is basically similar to the method embodiment, the description is relatively simple, and the relevant parts can be referred to the partial description of the method embodiment.

[0155] The steps in the methods of each embodiment of the present application can be adjusted in order, combined and deleted according to actual needs, and the technical features recorded in each embodiment can be replaced or combined.

[0156] The modules and sub-modules in the devices and terminals in the various embodiments of the present application can be combined, divided and deleted according to actual needs.

[0157] In the several embodiments provided in the present application, it should be understood that the disclosed terminals, devices and methods can be implemented in other ways. For example, the terminal embodiments described above are only schematic, for example, the division of modules or submodules is only a logical function division, and there may be other division methods in actual implementation, for example, multiple submodules or modules can be combined or integrated into another module, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be an indirect coupling or communication connection through some interfaces, devices or modules, which can be electrical, mechanical or other forms.

[0158] The modules or submodules described as separate components may or may not be physically separated, and the components of the modules or submodules may or may not be physical modules or submodules, that is, they may be located in one place, or they may be distributed on multiple network modules or submodules. Some or all of the modules or submodules may be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0159] In addition, each functional module or submodule in each embodiment of the present application may be integrated into one processing module, or each module or submodule may exist physically separately, or two or more modules or submodules may be integrated into one module. The above-mentioned integrated modules or submodules may be implemented in the form of hardware or in the form of software functional modules or submodules.

[0160] Professionals may further appreciate that the units and algorithm steps of each example described in conjunction with the embodiments disclosed herein can be implemented in electronic hardware, computer software, or a combination of the two. In order to clearly illustrate the interchangeability of hardware and software, the composition and steps of each example have been generally described in the above description according to function. Whether these functions are performed in hardware or software depends on the specific application and design constraints of the technical solution. Professionals and technicians may use different methods to implement the described functions for each specific application, but such implementation should not be considered to be beyond the scope of this application.

[0161] The steps of the method or algorithm described in conjunction with the embodiments disclosed herein may be implemented directly by hardware, software units executed by a processor, or a combination of the two. The software units may be placed in a random access memory (RAM), a memory, a read-only memory (ROM), an electrically programmable ROM, an electrically erasable programmable ROM, a register, a hard disk, a removable disk, a CD-ROM, or any other form of storage medium known in the art.

[0162] Finally, it should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the presence of other identical elements in the process, method, article or device including the elements.

[0163] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to the embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A spatial transepithelial impedance detection system, characterized in that: include: A cell culture pool and a detector; the cell culture pool comprises: a culture vessel, an upper electrode, a lower electrode array, and a thin film, wherein the thin film separates the culture vessel into an upper culture vessel and a lower culture vessel, wherein the upper culture vessel is used to grow epithelial cells above the thin film; one end of the upper electrode is inserted into the upper culture fluid in the upper culture vessel, and the other end is connected to the detector; the lower electrode array is located at the bottom of the lower culture vessel and is connected to the detector through a metal lead, wherein the height of the lower culture fluid in the lower culture vessel is at least greater than or equal to the height of the thin film; a circuit channel is formed between each lower electrode in the lower electrode array and the upper electrode; The detector is used to obtain the background impedance of each of the circuit channels when the epithelial cells are not grown in the cell culture tank; obtain the first transverse equivalent impedance of the transverse equivalent circuit of adjacent lower electrodes of the lower electrode array in the lower culture fluid; and, when the epithelial cells are grown in the cell culture tank, obtain the total impedance of each of the circuit channels; determine the second transverse equivalent impedance of the circuit channel by combining the first transverse equivalent impedance and the position of the lower electrode corresponding to the circuit channel in the lower electrode array; and determine the spatial transepithelial impedance corresponding to each of the circuit channels based on the total impedance of each of the circuit channels, the background impedance and the second transverse equivalent impedance.

2. The system according to claim 1, characterized in that The step of obtaining the background impedance of each of the circuit channels when the epithelial cells are not grown in the cell culture pool comprises: When the epithelial cells are not grown in the cell culture tank, obtaining an EIS impedance spectrum of each circuit channel of the cell culture tank; Determine, according to the EIS impedance spectrum, the electrical double layer impedance of the electrode surface of the upper electrode and the lower electrode of each of the circuit channels, the impedance of the upper culture solution, the impedance of the film, and the longitudinal equivalent impedance of the lower culture solution corresponding to each of the circuit channels; The background impedance of each circuit channel is determined according to the electrical double layer impedance of the electrode surface, the impedance of the upper culture solution, the impedance of the film and the longitudinal equivalent impedance.

3. The system according to claim 2, characterized in that The background impedance of each of the circuit paths is expressed by the following formula: Z F =2*Z CPE +R up +R B +R M ; Among them, Z F represents the background impedance; Z CPE represents the electrical double layer impedance of the electrode surface; R up Represents the impedance of the upper culture solution; R M represents the impedance of the film; R B Denotes the longitudinal equivalent impedance.

4. The system according to claim 1, characterized in that The step of obtaining a first lateral equivalent impedance of a lateral equivalent circuit of adjacent lower electrodes of the lower electrode array in the lower culture solution comprises: The first lateral equivalent impedance is obtained by scanning the EIS impedance spectrum through adjacent circuit channels.

5. The system according to claim 1, characterized in that The determining the second lateral equivalent impedance of the circuit channel by combining the first lateral equivalent impedance and the position of the lower electrode corresponding to the circuit channel in the lower electrode array comprises: Determining the position of each of the lower electrodes in the lower electrode array; Obtaining impedance coefficients corresponding to any two lower electrodes according to positions of the lower electrodes in the lower electrode array; A second transverse equivalent impedance of the circuit path is determined based on the impedance coefficient and the first transverse equivalent impedance.

6. The system according to claim 1, characterized in that Determining the spatial transepithelial impedance corresponding to each of the circuit channels according to the total impedance of each of the circuit channels, the background impedance, and the second transverse equivalent impedance includes: Constructing a correlation between the transepithelial impedance and the total impedance of each circuit channel, the background impedance, and the second transverse equivalent impedance; Based on the association relationship, the spatial transepithelial impedance corresponding to each of the circuit channels is obtained.

7. The system according to claim 6, characterized in that The association relationship can be expressed by the following formula: C=B -1 ×A; Among them, Z UX Represents the total impedance of the Xth circuit channel; Z F represents the background impedance; Z X represents the spatial transepithelial impedance corresponding to the Xth circuit channel; λ N～M Indicates the intermediate coefficient; R N～M It represents the second lateral equivalent impedance corresponding to the circuit channel composed of the lower electrode in the Nth row and the Mth column in the lower electrode array and the upper electrode; N and M are positive integers less than X.

8. The system according to claim 7, characterized in that R N～M Obtained by the following formula: R N～M =K N～M ×R0; Among them, K N～M represents the impedance coefficient corresponding to the Nth lower electrode to the Mth lower electrode; R0 represents the first lateral equivalent impedance.

9. A method for detecting spatial transepithelial impedance, characterized in that: The spatial transepithelial impedance detection system applied to any one of claims 1 to 8 above comprises: In the case where no epithelial cells are grown in the cell culture tank, the background impedance of each circuit channel of the cell culture tank is obtained; and the first lateral equivalent impedance of the lateral equivalent circuit of the adjacent lower electrodes of the lower electrode array of the cell culture tank in the lower culture fluid is obtained; When the epithelial cells are grown in the cell culture tank, obtaining the total impedance of each of the circuit channels; Determine, in combination with the first lateral equivalent impedance and the circuit structure of the lateral equivalent circuit, a second lateral equivalent impedance of any two lower electrodes in the lower electrode array in the lateral equivalent circuit; The spatial transepithelial impedance corresponding to each of the circuit channels is determined according to the total impedance of each of the circuit channels, the background impedance and the second transverse equivalent impedance.

10. An electronic device, characterized in that: include: processor; a memory for storing instructions executable by the processor; The processor is used to execute the spatial transepithelial impedance detection method of claim 9 by running instructions in the memory.

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