CN113652355A - Microfluidic device for electrical impedance monitoring and method for detecting cell electrical impedance - Google Patents
Microfluidic device for electrical impedance monitoring and method for detecting cell electrical impedance Download PDFInfo
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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Abstract
The invention provides a micro-fluidic device for electrical impedance monitoring and a method for detecting cell electrical impedance. The method comprises the following steps: the chip body consists of an upper cover body and a lower cover body, and the bottom of the upper cover body and the top of the lower cover body are provided with a microflow channel and a cavity communicated with the microflow channel; the cell culture chamber is arranged in the chip body and comprises interdigital electrodes arranged at the bottom in the chamber, parting strips among the interdigital electrodes, a cell culture carrier arranged in the chamber and auxiliary electrodes arranged at the top in the chamber; the cell culture carrier can be fixed on the upper part of the interdigital electrode parting strip or can be an independent plug-in, and the cell culture carrier can be an artificial cell basement membrane. The invention has the beneficial effects that: the real-time electrical impedance detection can be carried out on the cells cultured on the basement membrane, and the growth condition of the cells and the reaction of the medicine are analyzed through electrical impedance data; the device has the advantages of simple manufacturing process, simple manufacturing of the device, compact structure, low cost and batch production, and can be used for drug screening.
Description
Technical Field
The invention belongs to the technical field of biological detection, and particularly relates to a micro-fluidic device for electrical impedance monitoring and a method for detecting cell electrical impedance.
Background
Membrane-cultured cells or molecules have irreplaceable effects on biomimetic, drug screening assays. Biological based assays, such as those using labels, are based on end-point detection and have some effect on the cells. Electrophysiological study electrical methods one of the important methods for studying cells or molecules is cellular electrical impedance sensing.
The electrical impedance measurement technology is based on the system to correspondingly detect the physical, chemical and biological characteristics of a measured object by an alternating electric field, and the key of the electrical impedance measurement technology is related to the charge transfer generated on the contact surface of different substances. Typical electrical impedance measurements employ interdigitated electrodes. Typically interdigitated electrodes are fabricated on a flat surface and the cells are then detected on the electrodes and in the area between the electrodes. Due to the narrow gap between interdigitated electrodes, the molecules enriched using this design are limited and, if used for cell production, cell throughput is also limited. In order to solve the problem, a subsequent researcher adopts a culture chamber to be connected with the interdigital electrode, so that cells are cultured in the culture chamber, but the design can only test the cells cultured in the chamber adherent. In both designs, electrical impedance measurement cannot be carried out when cells or molecules are cultured in a membrane, the plane of the electric field intensity between the two electrodes is the strongest, the electric field intensity of the rest space distribution is the second highest, and the plane of the cell growth is different due to the uneven distribution of the electric field, so that the measurement sensitivity to the plane of the interdigital electrode is lower when the interdigital electrode is used. Meanwhile, the conventional design inserts the auxiliary electrode into the system, and the test result is also degraded due to operation error because of the inability of fixing the position.
Disclosure of Invention
In order to solve the technical problems, the invention provides a micro-fluidic device for electrical impedance monitoring and a method for detecting cell electrical impedance.
The specific technical scheme is as follows:
a microfluidic device for electrical impedance monitoring, characterized by comprising:
the chip comprises a chip body, wherein an inlet and an outlet are formed in the surface of the chip body;
the culture chamber is communicated with the opening and the inlet and is arranged in the chip body, and comprises a culture chamber inner bottom surface and a culture chamber inner upper surface which is arranged opposite to the culture chamber bottom surface;
the interdigital electrode is arranged in the chip body and comprises an interdigital electrode testing part and an interdigital electrode connecting part, and the interdigital electrode testing part is contacted with the inner space of the culture chamber;
the auxiliary electrode is arranged in the culture chamber;
the interlayer is arranged in the culture chamber and consists of a plurality of parting beads arranged between the interdigital electrodes and the auxiliary electrodes, one end of each parting bead is inserted between the adjacent positive electrode and the adjacent negative electrode of the interdigital electrode, and a space is reserved between the other end of each parting bead and the auxiliary electrode;
and the cell culture carrier is arranged in the culture chamber and is arranged between the interlayer and the auxiliary electrode.
Further, the cell culture carrier is a porous material or a cell culture membrane or a permeable membrane or a substance containing a permeable membrane.
Further, the cell culture carrier is connected with the other end of the parting strip.
Further, the cell culture carrier is a porous material or a permeable membrane or a substance containing a permeable membrane, and the cell culture carrier and the auxiliary electrode form a space.
Further, the chip body comprises an upper cover body and a lower cover body which are detachably spliced, the upper cover body comprises the inner upper surface of the culture chamber, and the lower cover body comprises the inner bottom surface of the cavity; the culture cavity further comprises an upper cover body cavity groove and a lower cover body cavity groove, and the upper cover body cavity groove and the lower cover body cavity groove can be spliced to form the culture cavity; the upper cover body cavity groove is formed in the inner bottom surface of the cavity, and the lower cover body cavity groove is formed in the inner bottom surface of the cavity; the inlet comprises a first inlet and a second inlet, the outlet comprises a first outlet and a second outlet, the first inlet and the first outlet are communicated with the upper cover body cavity groove through a microfluidic channel, and the second inlet and the second outlet are communicated with the lower cover body cavity groove through a microfluidic channel.
Furthermore, the auxiliary electrode is arranged inside the lower cover body cavity groove or inside the upper cover body cavity groove; the interdigital electrode is arranged inside the upper cover body cavity groove or inside the lower cover body cavity groove; the auxiliary electrode or the interdigital electrode is not arranged in the upper cover body cavity groove at the same time or in the lower cover body cavity groove at the same time.
Further, the cell culture carrier is disposed between the upper cover and the lower cover.
Further, the inlet and the outlet are connected with the culture chamber through a microfluidic channel; the micro-fluidic device for the electrical impedance detection further comprises a Lu-Lu.
Further, an upper cover body sealing layer is connected to the position, except the position where the upper cover body cavity groove is formed, of the upper surface in the culture cavity, and a lower cover body sealing layer is connected to the position, except the position where the lower cover body cavity groove is formed, of the inner bottom surface in the culture cavity.
The interdigital electrode is characterized by further comprising an external connection part connected with the chip body, wherein the external connection part is arranged at a position close to the interdigital electrode, and the interdigital electrode connection part is connected with the external connection part; and an auxiliary electrode connecting port is also formed on the surface of the chip body, which is close to the auxiliary electrode, and the auxiliary electrode connecting port is connected with the auxiliary electrode through a connecting channel.
Further, the lower cover body comprises a substrate and a lower cover body which are detachably connected, and the substrate comprises an inner bottom surface of the cavity; the microfluidic device for monitoring the electrical impedance further comprises an external connection part connected with the substrate, and the interdigital electrode connection part is connected with the external connection part.
Further, the inlet comprises a first inlet and a second inlet, the outlet comprises a first outlet and a second outlet, the first inlet and the first outlet are connected through a microfluidic channel, and the second outlet are connected through a microfluidic channel.
A method for testing the electrical impedance of cells by using the microporous flow device is characterized in that the cells are planted on the cell culture carrier, the adjacent positive electrode and the adjacent negative electrode of the interdigital electrode are communicated, and the electrical impedance is measured; and/or communicating the anode of the interdigital electrode with the auxiliary electrode to measure the electrical impedance; or the cathode of the interdigital electrode is communicated with the auxiliary electrode to measure the electrical impedance.
Compared with the prior art, the invention has the beneficial effects that:
(1) the interdigital electrode is isolated from the cell culture carrier by the interlayer with the parting strips, so that the electrical impedance detection can be carried out when the cells are cultured by the membrane without influencing the growth of the cells, the cells can be cultured on the whole cell culture carrier, the cell production is improved, and the cell growth condition is monitored in real time through electrical impedance data; the electric field generated by the interdigital electrode can reach the cell culture carrier without obstacles through the upper part of the parting strip, and is uniformly distributed in the cell culture carrier region, so that cell data growing on the cell culture carrier is acquired, and the detection sensitivity is improved; the auxiliary electrode is also integrated in the chip body, so that the subsequent operation error caused by moving the measuring position can be avoided, and the accuracy is improved; the electrical impedance of the whole growth of the cells can be obtained, and the electrical impedance data of the cells growing along the longitudinal direction of the cell culture carrier and penetrating the cell culture carrier can be detected, so that the three-dimensional detection is realized; the growth of the cells and the response of the drug were analyzed by impedance data.
(2) The invention adopts a split structure of the upper cover body and the lower cover body, and is convenient for disassembly operation.
(3) The auxiliary electrode and the interdigital electrode are respectively integrated on the upper cover body and the lower cover body, so that different materials can be adopted for preparation, and the process difficulty is reduced.
(4) The auxiliary electrode and the interdigital electrode are respectively arranged on the upper cover body and the substrate, so that subsequent replacement and splicing are convenient, the process difficulty is further reduced, and batch production is facilitated.
(5) The upper cover body cavity groove and the lower cover body cavity groove are connected through different micro-flow channels, the cell culture carrier can completely isolate two different spaces from the culture cavity, and the cell culture carrier capable of planting cells or integrating molecules on two sides can be designed, so that more complex experiments can be carried out.
(6) The cell culture carrier is a porous material or a cell culture membrane or a permeable membrane or a substance containing the permeable membrane, is arranged between the upper cover body and the lower cover body, and can detect the osmosis by using an electric impedance test.
Drawings
Fig. 1 is a cross-sectional view of a microfluidic device;
FIG. 2 is a microscopic top view of the cell culture carrier, the interlayer and the interdigital electrode after being connected;
FIG. 3 is a microscopic cross-sectional view A of FIG. 2;
fig. 4 is a cross-sectional view of the microfluidic device of example 2;
FIG. 5 is a schematic diagram of the electrical impedance measurement of the membrane cell culture in example 3;
FIG. 6 is a graph showing the results of electrical impedance measurement in the membrane cell culture of example 3.
The cell culture medium comprises a chip body-1, an inlet-101, an outlet-102, an upper cover body-103, a lower cover body-104, an auxiliary electrode connecting port-105, a substrate-1041, a lower cover body-1042, a culture chamber-2, a cavity inner bottom surface-2 a, an upper surface-2 b in the culture chamber, an upper cover body cavity groove-201, a lower cover body cavity groove-202, an interdigital electrode-3, an interdigital electrode testing part-301, an interdigital electrode connecting part-302, an interlayer-4, a parting bead-401, a cell culture carrier-5, an external part-6, a microfluidic channel-7, an auxiliary electrode-8, a connecting channel-9, a Ludao inlet-10, a Ludao outlet-11, an upper cover body sealing layer-12 and a lower cover body sealing layer-13.
Detailed Description
The present invention is further described in detail below with reference to specific examples so that those skilled in the art can more clearly understand the present invention.
The following examples are provided only for illustrating the present invention and are not intended to limit the scope of the present invention. All other embodiments obtained by a person skilled in the art based on the specific embodiments of the present invention without any inventive step are within the scope of the present invention.
In the examples of the present invention, all the raw material components are commercially available products well known to those skilled in the art, unless otherwise specified; in the examples of the present invention, unless otherwise specified, all technical means used are conventional means well known to those skilled in the art.
Example 1
The embodiment provides a microfluidic device for electrical impedance monitoring, the specific structure of which is shown in fig. 1, and the microfluidic device comprises:
the chip comprises a chip body 1, wherein an inlet 101 and an outlet 102 are arranged on the surface of the chip body 1. The culture cavity 2 is used for placing a sample or culture solution, the culture cavity 2 is arranged in the chip body 1 and is communicated with the inlet 101 and the outlet 102, and the culture cavity 2 comprises a culture cavity inner bottom surface 2a and a culture cavity inner upper surface 2b which is arranged opposite to the culture cavity inner bottom surface 2 a.
The interdigital electrode 3 comprises an interdigital electrode testing part 301 and an interdigital electrode connecting part 302, the interdigital electrode testing part 301 is in contact with the inner space of the culture chamber 2 and is used for detecting samples in the culture chamber 2 subsequently, the interdigital electrode connecting part 301 is connected with an external testing instrument, in the embodiment, the interdigital electrode 3 is designed by adopting a periodic structure with crossed anodes and cathodes, which is common in the field, the width between the adjacent anodes and cathodes is 10-100 mu m, and the width between each anode and cathode is 10-100 mu m, and the interdigital electrode is an Au electrode.
The isolation layer 4 and the cell culture carrier 5 are arranged in the culture chamber 2, the isolation layer 4 comprises a plurality of division bars 401, one ends of the division bars 401 are arranged in gaps of the interdigital electrodes 3, the other ends of the division bars 401 are connected with the cell culture carrier 5, in the embodiment, the division bars 401 are made of SU8, and the height (the thickness of the isolation layer) of the division bars 401 is 10-50 μm.
To reduce the error of the repeated experiments, the auxiliary electrodes 8 are arranged inside the culture space with a space between the cell culture carriers.
For the convenience of subsequent operations, the external connection portion 6 connected to the chip body 1 is provided at a position close to the interdigital electrode connection portion 302, and then connected to the interdigital electrode connection portion 302.
The cell culture carrier 5 is a porous material or a cell culture membrane or a permeable membrane or a substance containing a permeable membrane, and if the cell culture carrier is a cell culture membrane or a porous material capable of cell culture, the cell culture carrier can be used for cell adherent culture, in this embodiment, the cell culture carrier 5 is a nanowire mesh made of gelatin, and microscopic test charts of the interdigital electrode 3, the interlayer 4 and the cell culture carrier 4 are shown in fig. 2 to fig. 3.
In order to reduce the difficulty of the process integration manufacturing of the microfluidic chip, the chip body 1 is designed into a split structure in this embodiment, and specifically includes an upper cover 103 and a lower cover 104 which are detachably spliced, where the upper cover 104 includes an upper surface 2b in the culture chamber, and the lower cover 104 includes a bottom surface 2a in the cavity; the culture cavity 2 comprises an upper cover body cavity groove 201 and a lower cover body cavity groove 201, and the upper cover body cavity groove 201 and the lower cover body cavity groove 202 can be spliced into the culture cavity; the upper cover body cavity groove 201 is arranged on the inner upper surface 2b of the cavity, and the lower cover body cavity groove 202 is arranged on the inner bottom surface 2a of the cavity.
Meanwhile, in order to meet the experimental requirements of double-sided planting or research on permeation exchange effect, the device of the embodiment of the cell culture carrier can also be provided with two inlets 101 which are respectively a first inlet and a second inlet on the upper surface of the chip body 1; and two outlets 102, a first outlet and a second outlet, respectively. The upper cover body cavity groove 201 of the first inlet 101 is communicated with the first outlet 102 through a microfluidic channel; the second inlet 101, the lower cover body cavity groove 202 and the second opening 102 are communicated through the microfluidic channel 7. Two ends of the upper cover body cavity groove 201 are respectively provided with an opening which is connected with the micro-flow channel 7; two ends of the lower cover body cavity groove 202 are respectively provided with an opening which is connected with the microfluidic channel 7, and if cell culture is carried out on one surface of the cell culture carrier or molecules are integrated, culture solution can be filled in the upper cover body cavity groove 201; if the cell culture carrier 5 is used for double-sided cell culture or molecule integration, different culture solutions can be introduced into the upper cover body cavity groove 201 and the lower cover body cavity groove 202, and if the cell culture carrier 5 is a culture membrane which has permeability and can be used for double-sided cell planting, the study of complex interaction with permeability, such as the construction of blood brain barrier, can be carried out.
In order to facilitate the auxiliary electrodes 8 and the interdigital electrodes 3 to be subjected to different material replacement, chemical modification and other treatments according to the actual situation of use, the auxiliary electrodes 8 and the interdigital electrodes 3 are not designed inside the upper cap body cavity groove 201 and the lower cap body cavity groove 202 at the same time, but are designed separately. In this embodiment, the auxiliary electrode is a Pt electrode, the auxiliary electrode 8 is disposed inside the upper lid body cavity groove 201, in this embodiment, the auxiliary electrode 8 is connected to the upper surface 2b in the culture chamber, the auxiliary electrode connection port 105 corresponding to the auxiliary electrode is opened on the surface of the chip body 1, and the auxiliary electrode connection port 105 and the auxiliary electrode 8 are communicated by the connection channel 9; the interdigital electrode testing part 3, the interlayer 4 and the cell culture carrier 4 are all arranged inside the lower cover body cavity groove 202, specifically, the interdigital electrode testing part 301 is connected with the inner bottom surface 2a of the culture cavity, the interdigital electrode external connection part 302 penetrates through the chip body, and the tail end of the interdigital electrode external connection part is connected with the external connection part 6.
In order to further facilitate replacement and reduce the process difficulty, the lower cover body 104 is further split, specifically, the lower cover body 104 is split into the substrate 1041 and the lower cover body 1042 which are sequentially overlapped, the bottom surface 2a in the cavity is on the substrate, and the interdigital electrode test part 301 is connected with the bottom surface 2a in the cavity; in the present embodiment, the external portion 6 is integrally designed with the base 1041 at a position where the base 1041 extends relative to the lower cover body 1042.
In this embodiment, the chip body 1 is connected with the robust inlet 10 and the robust outlet 11, the robust inlet 10 is connected with the chip body 1 at a position where the inlet 101 is opened, and the robust outlet 11 is connected with the chip body 1 at a position where the outlet 102 is opened.
In this embodiment, in order to improve the sealing performance between the upper cover 103 and the lower cover 104, the upper cover sealing layer 12 is connected to the upper surface 2b of the culture chamber except the position where the upper cover cavity groove 201 is formed, the lower cover sealing layer 13 is connected to the inner bottom surface 2a of the culture chamber except the position where the lower cover cavity groove 202 is formed, and both the upper cover sealing layer 12 and the lower cover sealing layer 13 are made of soft materials.
Example 2
The embodiment provides a microfluidic device for electrical impedance monitoring, and the specific structure is shown in fig. 4, and includes:
the chip body, the chip body is including dismantling the upper cover body 103 and the lower cover body 104 of concatenation, upper cover body 104 is including cultivateing indoor higher authority 2b of cavity, lower cover body 104 is including superpose basement 1041 and lower cover body 1042 in proper order, the basement includes upper cover body cavity inner bottom surface 2a, upper cover body cavity groove 201 has been seted up to the indoor higher authority of cultivation cavity, lower cover body cavity groove 202 has been seted up to bottom surface 2a in the cavity, upper cover body cavity groove 201 and lower cover body cavity groove 202 can splice into the cultivation cavity, the position that upper cover body cavity groove 201 was seted up except to cultivate indoor higher authority 2b of cavity is connected with upper cover body sealing layer 12, the position that lower cover body cavity groove 202 was set up to bottom surface 2a in the cultivation cavity is divided to open is connected with lower cover body sealing layer 13, upper cover body sealing layer 12 and lower cover body sealing layer 13 all adopt soft material to prepare.
An external connection portion 6 connected to the chip body, the external connection portion 6 and the substrate 1041 are integrally designed, and are located at a position where the substrate 1041 extends relative to the lower cover body 1042.
The upper surface of the chip body 1 is simultaneously provided with two inlets 101 which are respectively a first inlet and a second inlet; and two outlets 102, a first outlet and a second outlet, respectively. The first inlet 101, the upper cover body cavity groove 201 and the first outlet 102 are communicated through the microfluidic channel 7; the second inlet 101, the lower cover body cavity groove 202 and the second opening 102 are communicated through the microfluidic channel 7. Specifically, two ends of the upper cover body cavity groove 201 are respectively provided with an opening which is connected with the micro-flow channel 7; two ends of the lower cover body cavity groove 202 are respectively provided with an opening which is connected with the micro-flow channel 7.
The interdigital electrode 3 is connected with the chip body, the interdigital electrode comprises an interdigital electrode testing part 301 and an interdigital electrode external connection part 302, the interdigital electrode testing part 301 is connected with the inner bottom surface 2a of the cavity at the cavity groove 202 of the lower cover body, the interdigital electrode external connection part 302 penetrates through the chip body, and the tail end of the interdigital electrode external connection part is connected with the external connection part.
And the interlayer 4 consists of a plurality of parting beads 401, one end of each parting bead is inserted between the adjacent positive electrode and negative electrode gaps of the interdigital electrode 3, the other end of each parting bead is arranged in the lower cover body cavity groove 202, and the height of the whole interlayer does not exceed the depth of the lower cover body cavity groove 202.
And the cell culture carrier 5 is arranged between the upper cover body 103 and the lower cover body 104, the cell culture carrier is a porous material or a permeable membrane or a substance containing the permeable membrane, and the cell culture carrier 5 can be fixed after the upper cover body 103 is connected with the lower cover body 104.
The auxiliary electrode 8 is connected to the upper surface 2b of the culture chamber in the cavity groove 202 of the upper lid, and an auxiliary electrode connection port 105 is formed on the surface of the chip body 1 to correspond to the auxiliary electrode connection port 105, and the auxiliary electrode connection port 105 and the auxiliary electrode 8 are connected by a connection channel 9.
Example 3
In this example, to further verify the feasibility of the chip in example 1 in monitoring cell growth, the following verification tests were performed:
(1) when the nano-mesh was not laid, medium (cell culture solution) was added to the chip, and a second generation Analog Discovery analyzer (Analog Discovery 2) of Analog Devices, USA, was usedTM) The medium was tested for impedance spectroscopy with a test voltage of 200mV and a frequency of 102～106Hz, communicating the adjacent positive electrode and the adjacent negative electrode of the interdigital electrode in the test of the impedance spectrum, and performing three times of parallel electrical Impedance (IDE) of the adjacent positive electrode and the adjacent negative electrode, wherein the electric field is the electric field between the positive electrode and the adjacent interdigital electrode as shown in figure 5; and then communicating the anode of the interdigital electrode with the auxiliary electrode or communicating the cathode of the interdigital electrode with the auxiliary electrode to perform three times of parallel electrical Impedance (ICE) tests of gaps between the adjacent anode and cathode of the interdigital electrode, as shown in fig. 5, wherein the electric field is an electric field between the anode and the auxiliary electrode or an electric field between the auxiliary electrode and the cathode. Obtaining the medium, ICE with nanofibers and medium in FIG. 6,IDE without nanofibers curve.
(2) And cleaning the interdigital electrode after the test is finished, and depositing a layer of nano-wire mesh on the interdigital electrode by utilizing an electrostatic spinning technology. The interdigital electrode with the nanofiber deposited on the surface is subjected to IDE and ICE tests of the medium in the same way as the operation, and medium, ICE with nanofibers and medium, IDE with nanofibers curves in figure 6 are obtained.
(3) After the completion of the test, SK-OV-3 cells were seeded on nanofibers, and after one hour of culture in an incubator, IDE and ICE tests were performed on interdigital electrodes with nanofibers deposited on the surface and cells seeded thereon in the same manner as described above, to obtain the curves for cells, ICE with nanofibers and cells, IDE with nanofibers in FIG. 6.
The results show that the impedance of the interdigitated electrodes with nanofibers is higher than without nanofibers but lower than with cells. In the presence of nanofibers (or cells), the impedance of the IDE test is generally higher than that of the ICE test because the current between the interdigitated electrodes must cross the nanofibers (or cell layers) above the positive electrode and the nanofibers (or cell layers) above the negative electrode when the IDE test is performed, whereas the current between the positive or negative electrode of the interdigitated electrodes and the auxiliary electrode only needs to cross one layer of nanofibers (or cell layers) above the positive electrode or one layer of nanofibers (or cell layers) above the negative electrode when the ICE test is performed. The impedance of the IDE test may be lower than that of the ICE test if no nanofibers (or cells) are present, depending on the effective distance between participating electrodes and the nature of the solution-electrode interface.
Based on the above, IDE data can reflect the overall growth condition of cells, ICE data can reflect the overall growth condition of cells to a certain extent, and data of cells growing longitudinally along a cell culture carrier can be fed back, so that the data provides a good basis for subsequent researches on barrier construction, permeability and the like.
It should be noted that the above examples are only for further illustration and description of the technical solution of the present invention, and are not intended to further limit the technical solution of the present invention, and the method of the present invention is only a preferred embodiment, and is not intended to limit the protection scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A microfluidic device for electrical impedance monitoring, comprising:
the chip comprises a chip body, wherein an inlet and an outlet are formed in the surface of the chip body;
the culture chamber is communicated with the opening and the inlet and is arranged in the chip body, and comprises a culture chamber inner bottom surface and a culture chamber inner upper surface which is arranged opposite to the culture chamber bottom surface;
the interdigital electrode is arranged in the chip body and comprises an interdigital electrode testing part and an interdigital electrode connecting part, and the interdigital electrode testing part is contacted with the inner space of the culture chamber;
the auxiliary electrode is arranged in the culture chamber;
the interlayer is arranged in the culture chamber and consists of a plurality of parting beads arranged between the interdigital electrodes and the auxiliary electrodes, one end of each parting bead is inserted between the adjacent positive electrode and the adjacent negative electrode of the interdigital electrode, and a space is reserved between the other end of each parting bead and the auxiliary electrode;
and the cell culture carrier is arranged in the culture chamber and is arranged between the interlayer and the auxiliary electrode.
2. The microfluidic device for electrical impedance monitoring according to claim 1, wherein the cell culture carrier is a porous material or a cell culture membrane or a permeable membrane or a substance containing a permeable membrane.
3. The microfluidic device for electrical impedance monitoring of claim 1, wherein the cell culture carrier is connected to the other end of the spacer.
4. The microfluidic device for electrical impedance monitoring according to claim 1, wherein the cell culture carrier is a porous material or a permeable membrane or a substance containing a permeable membrane, and a space exists between the cell culture carrier and the auxiliary electrode.
5. The microfluidic device for electrical impedance monitoring according to claim 1 or 2, wherein the chip body comprises an upper cover body and a lower cover body which are detachably spliced, the upper cover body comprises an upper surface in the culture chamber, and the lower cover body comprises a bottom surface in the cavity; the culture cavity further comprises an upper cover body cavity groove and a lower cover body cavity groove, and the upper cover body cavity groove and the lower cover body cavity groove can be spliced to form the culture cavity; the upper cover body cavity groove is formed in the inner bottom surface of the cavity, and the lower cover body cavity groove is formed in the inner bottom surface of the cavity; the inlet comprises a first inlet and a second inlet, the outlet comprises a first outlet and a second outlet, the first inlet and the first outlet are communicated with the upper cover body cavity groove through a microfluidic channel, and the second inlet and the second outlet are communicated with the lower cover body cavity groove through a microfluidic channel.
6. The microfluidic device for electrical impedance monitoring of claim 3, wherein the auxiliary electrode is disposed inside the lower cover cavity slot or inside the upper cover cavity slot; the interdigital electrode is arranged inside the upper cover body cavity groove or inside the lower cover body cavity groove; the auxiliary electrode or the interdigital electrode is not arranged in the upper cover body cavity groove at the same time or in the lower cover body cavity groove at the same time.
7. The microfluidic device for electrical impedance monitoring of claim 6, wherein the cell culture carrier is disposed between the upper cover and the lower cover.
8. The microfluidic device for electrical impedance tomography monitoring according to claim 1, further comprising an external connection portion connected to the chip body, the external connection portion being disposed at a position close to the interdigital electrode, the interdigital electrode connection portion being connected to the external connection portion; and an auxiliary electrode connecting port is also formed on the surface of the chip body, which is close to the auxiliary electrode, and the auxiliary electrode connecting port is connected with the auxiliary electrode through a connecting channel.
9. The microfluidic device for electrical impedance monitoring of claim 3 or 4, wherein the lower cover comprises a substrate and a lower cover body which are detachably connected, the substrate comprises the inner bottom surface of the cavity; the microfluidic device for monitoring the electrical impedance further comprises an external connection part connected with the substrate, and the interdigital electrode connection part is connected with the external connection part.
10. A method for testing the electrical impedance of cells by using the microporous flow device as claimed in any one of claims 1 to 9, wherein the cells are planted on the cell culture carrier, the adjacent positive electrode and the adjacent negative electrode of the interdigital electrode are communicated, and the electrical impedance is measured; and/or communicating the anode of the interdigital electrode with the auxiliary electrode to measure the electrical impedance; or the cathode of the interdigital electrode is communicated with the auxiliary electrode to measure the electrical impedance.
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