Array chip integrating impedance measurement and microscopic observation

Abstract

The utility model relates to the technical field of bioengineering, and provides an array chip integrating impedance measurement and microscopic observation, which comprises a transparent substrate, a circuit board, a 3D printing board and an electrode unit, and one side of the transparent substrate is provided with a conductive film; the circuit board is arranged on the conductive film, and a plurality of micro-channels are arranged on the circuit board in an array manner; the 3D printing plate is arranged on the side, away from the conductive film, of the circuit board, a plurality of micro-cavities are arranged on the 3D printing plate in an array mode, and the positions of the micro-cavities correspond to the positions of the micro-channels one to one; the electrode unit comprises a first electrode plate and a second electrode plate, the 3D printing plate is covered with the first electrode plate, the circuit board is surrounded by the second electrode plate, and the second electrode plate and the first electrode plate form a closed loop through a conductive film. Through innovative fusion of micromachining and bioelectrochemistry, synchronization of electrophysiology (impedance) and morphology (microscopy) is realized, and the dynamic growth process of organisms is conveniently and continuously monitored.

Description

Technical Field

[0001] This invention relates to the field of bioengineering technology, and in particular to an array chip that integrates impedance measurement and microscopic observation. Background Technology

[0002] Organoids, as a type of three-dimensional cellular structure cultured in vitro, are of great significance in biomedical research, especially in the field of tumor research, where tumor organoids can mimic the physiological and pathological characteristics of tumors in vivo.

[0003] After regulating gene expression or administering chemotherapy drugs, organoids need to be obtained at different time points for analysis. However, traditional methods for monitoring organoid growth have several shortcomings: for example, observing organoid growth using an optical microscope, or detecting organoid growth using immunofluorescence staining and chemical reagents. While observing organoid growth using an optical microscope is intuitive, it only provides morphological information, and the organoids are processed after the analysis and cannot be used for further culture. Immunofluorescence staining and chemical reagents can alter the characteristics of the organoids.

[0004] Therefore, neither imaging-based detection methods nor conventional methods such as fluorescence labeling can achieve continuous real-time monitoring of tumor organoids within a single culture well. This means that there is a lack of information in studying the dynamic growth of organoids, and the changes in organoids throughout their entire growth cycle cannot be fully reflected. This limits a deeper understanding of organoid growth mechanisms and their application in drug development, disease diagnosis, and other fields. Utility Model Content

[0005] This invention provides an array chip that integrates impedance measurement and microscopic observation to address the aforementioned technical deficiencies in the prior art. Through the innovative fusion of microfabrication and bioelectrochemistry, it achieves synchronization of electrophysiology (impedance) and morphology (microscopy), facilitating continuous monitoring of the dynamic growth process of organisms.

[0006] This invention provides an array chip integrating impedance measurement and microscopic observation, comprising:

[0007] A transparent substrate with a conductive film on one side;

[0008] A circuit board is disposed on the conductive film, and the circuit board has an array of multiple microchannels.

[0009] A 3D printing plate is disposed on the side of the circuit board away from the conductive film. The 3D printing plate has an array of multiple microcavities, and the positions of the microcavities correspond one-to-one with the positions of the microchannels.

[0010] Electrode unit, including:

[0011] A first electrode plate is placed on the 3D printed plate. The first electrode plate is in contact with the sample in the microchannel. The first electrode plate is adapted to be connected to an impedance measuring instrument for applying an excitation signal to perform impedance detection.

[0012] A second electrode plate is disposed around the circuit board, and the second electrode plate forms a closed circuit with the first electrode plate through the conductive film.

[0013] According to the integrated impedance measurement and microscopic observation array chip provided by this utility model, the transparent substrate and the circuit board are bonded and fixed together by a first UV adhesive layer.

[0014] And / or, the circuit board and the 3D printed board are bonded and fixed together by a second UV adhesive layer.

[0015] According to the integrated impedance measurement and microscopic observation array chip provided by this utility model, the transparent substrate includes any one of sodium-calcium glass and borosilicate glass.

[0016] According to the array chip for integrated impedance measurement and microscopic observation provided by this utility model, the conductive film includes an indium tin oxide film.

[0017] According to the integrated impedance measurement and microscopic observation array chip provided by this utility model, the circuit board includes a flame-retardant epoxy resin glass fiber laminate.

[0018] According to the array chip for integrated impedance measurement and microscopic observation provided by this invention, the 3D printed plate includes a biocompatible resin plate.

[0019] According to the integrated impedance measurement and microscopic observation array chip provided by this utility model, the first electrode plate is provided with an in-hole pin and a first pin; the position of the in-hole pin corresponds one-to-one with the position of the microcavity, the first pin is symmetrically arranged on both sides of the first electrode plate, the first pin is connected to the in-hole pin one-to-one, and the first pin is suitable for connection with an impedance measurement instrument.

[0020] The second electrode is connected to the conductive film of the transparent substrate by UV adhesive curing, and the second electrode has a second lead of the conductive film.

[0021] According to the integrated impedance measurement and microscopic observation array chip provided by this utility model, at least two limiting members are connected to the first electrode by UV adhesive curing, and each pair of limiting members is located at two opposite apex corners of the first electrode.

[0022] When the first electrode plate is placed on the 3D printing plate, the limiting member abuts against the two opposite sidewalls of the 3D printing plate.

[0023] According to the integrated impedance measurement and microscopic observation array chip provided by this utility model, the aperture of the microchannel is 0.4-1.0 mm, and the center-to-center distance between two adjacent microchannels is 9 mm.

[0024] According to the integrated impedance measurement and microscopic observation array chip provided by this utility model, the diameter of the microcavity is 4-6 mm, the height of the microcavity is 5.4 mm, and the center-to-center distance between two adjacent microcavities is 9 mm.

[0025] The integrated impedance measurement and microscopic observation array chip provided by this invention offers a high spatiotemporal precision integrated solution for cell analysis through the innovative fusion of microfabrication and bioelectrochemistry. It achieves synchronization of electrophysiology (impedance) and morphology (microscopy), facilitating continuous monitoring of the dynamic growth process of organisms.

[0026] Furthermore, multi-channel parallel detection improves efficiency compared to single-well plates while reducing reagent consumption. The ITO conductive film layer combines conductive pathways with optical transparency, replacing traditional leads and improving the signal-to-noise ratio. The detachable first electrode plate supports repeated sterilization (alcohol immersion / UV irradiation), reducing the cost per use. Attached Figure Description

[0027] To more clearly illustrate the technical solutions in this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this utility model. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0028] Figure 1 This is a schematic diagram of the structure of the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention.

[0029] Figure 2 This is a partial structural schematic diagram of the array chip for integrated impedance measurement and microscopic observation provided in an embodiment of this utility model.

[0030] Figure 3 This is a front view of the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention.

[0031] Figure 4 This is a schematic diagram of the first electrode plate in the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention.

[0032] Figure 5 This is a schematic diagram of the second electrode plate in the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention.

[0033] Figure 6 This is a graph showing the change in impedance of each channel of the array chip for integrated impedance measurement and microscopic observation as a function of cell line concentration, provided in this embodiment of the present invention.

[0034] Figure label:

[0035] 10. Transparent substrate; 20. Circuit board; 21. Microchannel; 30. 3D printing plate; 31. Microcavity; 40. Electrode unit; 41. First electrode plate; 411. In-hole pin; 412. First pin; 42. Second electrode plate; 421. Second pin; 43. Limiting component; 50. First UV adhesive layer; 60. Second UV adhesive layer. Detailed Implementation

[0036] To make the objectives, technical solutions, and advantages of this utility model clearer, the technical solutions of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this utility model, not all embodiments. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of this utility model.

[0037] In the description of the embodiments of this application, it should be noted that, unless otherwise explicitly specified and limited, the terms "connected" and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium. Those skilled in the art can understand the specific meaning of the above terms in the embodiments of this application based on the specific circumstances.

[0038] In the embodiments of this application, unless otherwise expressly specified and limited, "above" or "below" the second feature can mean that the first feature is in direct contact with the second feature, or that the first feature is in indirect contact with the second feature through an intermediate medium. Furthermore, "above," "on top of," and "over" the second feature can mean that the first feature is directly above or diagonally above the second feature, or simply that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature can mean that the first feature is directly below or diagonally below the second feature, or simply that the first feature is at a lower horizontal level than the second feature.

[0039] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the embodiments of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0040] Figure 1 This is a schematic diagram of the structure of the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention. Figure 2 This is a partial structural schematic diagram of the array chip for integrated impedance measurement and microscopic observation provided in an embodiment of this utility model.

[0041] See Figure 1 and Figure 2 This utility model provides an array chip that integrates impedance measurement and microscopic observation.

[0042] The integrated impedance measurement and microscopic observation array chip includes a transparent substrate 10, a circuit board 20, a 3D printed plate 30, and an electrode unit 40.

[0043] The transparent substrate 10 is made of a transparent material, such as a glass substrate composed of either soda-lime glass or borosilicate glass. An ITO (indium tin oxide) conductive film is sputtered onto the surface of the transparent substrate 10 using magnetron sputtering to form ITO conductive glass, that is, a conductive film (indium tin oxide film) is provided on one side of the transparent substrate 10.

[0044] In some embodiments, transparent conductive materials such as graphene or silver nanowires can also be used as the conductive film. Transparent materials such as PDMS (polydimethylsiloxane) or COC (cyclic olefin copolymer) can also be used as the transparent substrate 10, taking into account optical transparency, chemical stability, and cell culture requirements.

[0045] The circuit board 20 is disposed on a conductive film, and multiple microchannels 21 are arrayed on the circuit board 20. These microchannels 21 can be used for 3D cell culture or organoid culture. The pore size of the microchannels 21 is 0.4-1.0 mm, and the center-to-center distance between two adjacent microchannels 21 is 9 mm. The circuit board 20 is composed of a flame-retardant epoxy resin glass fiber laminate, namely FR-4 (Flame Retardant 4) material. This material is composed of an epoxy resin matrix and glass fiber cloth, and flame retardants (such as brominated epoxy resin) are added to meet flame retardant standards. The circuit board 20 is easy to drill, etch, and UV adhesive bonded (such as bonding with ITO conductive glass).

[0046] This invention is illustrated using a 4×4 array of 16 microchannels 21 on a circuit board 20 as an example. In addition, the circuit board 20 can also have a 6×8 array of 48 microchannels 21, etc. Each microchannel 21 is pre-coated with fibronectin to enhance cell adhesion.

[0047] The 3D printing plate 30 is located on the side of the circuit board 20 away from the conductive film. The 3D printing plate 30 has an array of microcavities 31, the positions of which correspond one-to-one with the positions of the microchannels 21. The 3D printing plate 30 can be made of biocompatible resin (medical resin), and a plate with a thickness of 5.4 mm is printed using micro-stereolithography. The diameter of each microcavity 31 is 4-6 mm, the height of each microcavity 31 is 5.4 mm, and the center-to-center distance between two adjacent microcavities 31 is 9 mm.

[0048] The electrode unit 40 includes a first electrode plate 41 and a second electrode plate 42. The first electrode plate 41 is disposed on the 3D printing plate 30 and is in contact with the sample in the microchannel 21. The first electrode plate 41 can be a gold thin film electrode and is connected to an external impedance measuring instrument through leads for applying an excitation signal to perform impedance detection.

[0049] The second electrode plate 42 can be a ring-shaped platinum electrode. The second electrode plate 42 is arranged around the edge of the circuit board 20. The second electrode plate 42 is fixedly connected to the conductive film. The second electrode plate 42 forms a closed circuit with the first electrode plate 41 through the ITO conductive film.

[0050] The application scenarios of the array chip for integrated impedance measurement and microscopic observation provided by this utility model are as follows:

[0051] When using the aforementioned integrated impedance measurement and microscopic observation array chip for cell culture, cell suspension is injected into microchannel 21 for uniform distribution via capillary action, and the conductive membrane is grounded to avoid electrostatic interference. Culture medium is pre-loaded into microcavities 31, and cross-contamination is prevented by the hydrophobic walls of the 3D-printed plate 30. After a certain culture period, an AC excitation signal is applied to create an equivalent circuit consisting of the first electrode, cell layer, and conductive membrane. The changes in cell impedance values ​​in each microchannel 21 are then monitored in real time. Changes in cell membrane integrity (such as cell death) cause an increase in impedance modulus, and the data is acquired in a time-division multiplexing manner via a multiplexer. The impedance data is analyzed in real time using LabVIEW to generate a cell index curve.

[0052] Meanwhile, the transparent substrate 10 allows for bright-field / fluorescence imaging using objectives (such as Calcein-AM / PI double staining to verify cell viability) to observe apoptotic features such as cell morphology shrinkage and shedding. Image analysis algorithms (such as OpenCV) automatically identify cell coverage area and movement trajectory.

[0053] When drug treatment is performed using the array chip integrating impedance measurement and microscopic observation, different concentrations of chemotherapeutic drugs (such as cisplatin) are injected into the corresponding microcavities 31, and the drugs diffuse into the microchannels 21. Then, the changes in cell impedance values ​​in each microchannel 21 are monitored in real time, and apoptotic characteristics such as cell morphology shrinkage and shedding are observed using an inverted microscope.

[0054] It is understood that the integrated impedance measurement and microscopic observation array chip provided by this utility model, through the innovative fusion of microfabrication and bioelectrochemistry, provides a high spatiotemporal precision integrated solution for cell analysis, realizing the synchronization of electrophysiology (impedance) and morphology (microscopy), which facilitates continuous monitoring of the dynamic growth process of organisms.

[0055] Furthermore, multi-channel parallel detection improves efficiency compared to single-well plates while reducing reagent consumption. The ITO conductive film layer combines conductive pathways with optical transparency, replacing traditional leads and improving the signal-to-noise ratio. The detachable first electrode plate 41 supports repeated sterilization (alcohol immersion / UV irradiation), reducing the cost per use.

[0056] Continue reading Figure 2 In some embodiments of this invention, the transparent substrate 10 and the circuit board 20 are bonded and fixed together by a first UV adhesive layer 50. That is, liquid UV adhesive is uniformly coated on one side of the circuit board 20, the adhesive residue in the microchannel 21 is removed with a needle, the side of the circuit board 20 coated with UV adhesive is aligned with the conductive film of the ITO guide glass, pressure is applied to press them together, and then a UV light source with a wavelength of 395nm is used to irradiate to form the first UV adhesive layer 50.

[0057] UV adhesive is applied to the 3D printing plate 30, and excess UV adhesive is removed from the holes using a needle. The microcavities 31 of the 3D printing plate 30 are aligned with the microchannels 21 of the circuit board 20, and pressure is applied for pre-fixation.

[0058] In addition, the transparent substrate 10 and the circuit board 20 can be bonded and fixed together by the first UV adhesive layer 50, or the circuit board 20 and the 3D printing board 30 can be bonded and fixed together by the second UV adhesive layer 60.

[0059] Figure 3 This is a front view of the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention. Figure 4 This is a schematic diagram of the first electrode plate in the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention. Figure 5 This is a schematic diagram of the second electrode plate in the array chip for integrated impedance measurement and microscopic observation provided in this embodiment of the present invention.

[0060] Continue reading Figure 1 And see also Figures 3 to 5 In some embodiments of this utility model, the first electrode plate 41 is provided with an internal pin 411 and a first pin 412; the position of the internal pin 411 corresponds one-to-one with the position of the microcavity 31, and the first pin 412 is symmetrically arranged on both sides of the first electrode plate 41, and the first pin 412 is connected to the internal pin 411 one-to-one, and the first pin 412 is suitable for connection with an impedance measuring instrument. The second electrode plate 42 is connected to the conductive film of the transparent substrate 10 by UV adhesive curing, and the second electrode plate 42 has a second pin 421 with conductive film leading out.

[0061] During the fabrication of the first electrode plate 41, photolithography can be used to determine the position of the pins 411 inside the hole, ensuring that the position of the pins 411 inside the hole corresponds one-to-one with the position of the microcavity 31. The first pins 412 can be formed using processes such as metal deposition and etching, and metal wires (such as wires made of materials with good conductivity like gold or silver) are used to connect the first pins 412 to the pins 411 inside the hole one-to-one, ensuring the stability of the connection and low resistance characteristics.

[0062] During the fabrication of the second electrode plate 42, the second electrode plate 42 and the conductive film are aligned and fixed with clips. UV glue is dripped into the small hole, cured by irradiation with ultraviolet light for 1 minute, and left to stand for 1 hour before the clips are removed, forming a firm and conductive connection. When leading out the second pin 421, a process similar to that used to create the pins on the first electrode plate 41 is employed, such as metal deposition and etching, to lead out the second pin 421 of the conductive film from the second electrode plate 42.

[0063] When impedance measurement is required, the first pin 412 is connected to the impedance measuring instrument. Since the first pin 412 is connected to the pin 411 inside the hole in a one-to-one correspondence, and the pin 411 inside the hole corresponds to the position of the microcavity 31 in a one-to-one correspondence, and the microcavity 31 corresponds to the position of the microchannel 21 in a one-to-one correspondence, an electrical path from the microchannel 21 to the impedance measuring instrument is established.

[0064] During the measurement process, any changes in electrical properties that may occur within the microchannel 21 (such as changes in capacitance and resistance) are conducted through the pin 411 inside the hole to the first pin 412, and then detected by the impedance measuring instrument. This accurately reflects the electrical properties within the microchannel 21 into the measurement circuit, thereby improving the accuracy of the measurement and helping to detect information such as the concentration or type of biomolecules.

[0065] The first pin 412 is symmetrically positioned on both sides of the first electrode plate 41 and securely connected. This arrangement helps reduce the impact of electromagnetic interference and other factors on signal transmission, enabling the signal to be stably transmitted from the microchannel 21 to the impedance measuring instrument. The second pin 421, extending from the second electrode, provides the system with more electrical connection possibilities. For example, other electronic components or circuit modules can be connected through this pin to achieve more functions, such as signal amplification and filtering, thereby expanding the functionality and application range of the entire device.

[0066] Continue reading Figure 1 In some embodiments of this invention, at least two limiting members 43 are connected to the first electrode by UV adhesive curing, with each pair of limiting members 43 located at two opposite apex corners of the first electrode. When the first electrode plate 41 covers the 3D printing plate 30, the limiting members 43 abut against two opposite sidewalls of the 3D printing plate 30.

[0067] The limiting component 43 can be made of a material with certain hardness and precision, such as plastic or ceramic. The limiting component 43 is placed at two opposite apex positions of the first electrode plate 41, and then cured and connected using UV adhesive.

[0068] When the first electrode plate 41 is placed on the 3D printing plate 30, ensure that the orientation of the first electrode plate 41 is correct. As the first electrode plate 41 gradually approaches the 3D printing plate 30, since the limiting member 43 is located at the two opposite apex corners of the first electrode plate 41, the limiting member 43 first contacts the two opposite sidewalls of the 3D printing plate 30, ensuring accurate alignment of the first electrode plate 41 and the 3D printing plate 30, so that the limiting member 43 can stably abut against the sidewall of the 3D printing plate 30.

[0069] Meanwhile, the limiting member 43 serves a positioning function. Because the limiting member 43 abuts against the two opposing sidewalls of the 3D printing plate 30, it restricts the horizontal movement of the first electrode plate 41, ensuring that the first electrode plate 41 can accurately cover the 3D printing plate 30 and guaranteeing their relative positional relationship. During use, the limiting member 43 continuously abuts against the sidewalls of the 3D printing plate 30, preventing the first electrode plate 41 from shifting when affected by external vibrations, interference, or internal stress.

[0070] It is understood that in this embodiment of the invention, the limiting member 43 ensures that the first electrode plate 41 can be accurately and quickly installed onto the 3D printing plate 30, improving assembly efficiency and accuracy. In mass production or equipment that requires frequent replacement of the first electrode plate 41, this design can reduce debugging time and labor costs during the installation process.

[0071] The limiting member 43 abuts against the side wall of the 3D printing plate 30, effectively enhancing the stability of the first electrode plate 41 on the 3D printing plate 30. This helps maintain the stability of the electrical connection or other functional connection between the first electrode plate 41 and the 3D printing plate 30. For example, if there is a signal transmission line between the first electrode plate 41 and the 3D printing plate 30, a stable connection can reduce interference and loss during signal transmission.

[0072] In some embodiments of this utility model, the limiting member 43 has a slot, which is engaged on the first electrode plate 41 to achieve proper positioning.

[0073] The assembly process of the integrated impedance measurement and microscopic observation array chip provided in this embodiment of the invention is as follows:

[0074] A layer of liquid UV adhesive (approximately 0.1 mm thick) is applied to one side of the circuit board 20. The UV adhesive in the microchannel 21 is removed using a pinhole. The side of the circuit board 20 coated with UV adhesive is then bonded to the conductive surface of the ITO conductive glass. The circuit board 20 and the ITO conductive glass are pressed together. With no gaps between the circuit board 20 and the ITO conductive glass and minimal adhesive overflow in the microchannel 21, the circuit board is cured by irradiating it with 395 nm ultraviolet light at a power of 7 W for one minute.

[0075] Liquid UV adhesive is also used to bond the other side of the circuit board 20 to the 3D printing board 30. The second electrode plate 42 is connected to the conductive surface of the ITO conductive glass by dripping liquid UV adhesive into the hole of the second electrode plate 42 and curing it with ultraviolet light. This allows the conductive film electrode of the ITO conductive glass to be led out to the third pin of the second electrode.

[0076] The limiting member 43 on the first electrode plate 41 is also bonded to the first electrode plate 41 with UV adhesive, so that when the first electrode plate 41 is placed on the 3D printing plate 30, the test probe is in the same position, ensuring the accuracy of impedance measurement.

[0077] In this embodiment of the invention, the overall size of the array chip is 60*60*6.5mm, and the size of the ITO conductive glass is 60*60*0.5mm. The main function of the ITO conductive glass is to provide a chip substrate and serve as one end of the impedance measurement electrode, while also facilitating microscopic observation. The size of the circuit board 20 is 50*50*0.4mm. The microchannels 21 set in the circuit board 20 are used as the main containers for cell line growth and organoid growth. The smaller the diameter of the microchannels 21, the higher the measurement accuracy, but the greater the manufacturing difficulty.

[0078] The 3D printed plate 30 is used to hold the culture medium required for the growth of cell lines and organoids. The height of the 3D printed plate 30 is 5.4 mm, which allows the pins of the first electrode plate 41 to extend closer to the circuit board 20, thereby improving the accuracy of impedance measurement. The main function of the second electrode plate 42 is to lead out a pin header from the conductive film on the ITO conductive glass.

[0079] During use, the cell suspension is dripped into the well using a pipette, and the cell image can be observed under a microscope. The first electrode plate 41 is then covered, and the impedance value of each microchannel 21 can be measured by connecting the first pin 412 and the second pin 421, which are electrically connected to the pin 411 inside the well, using an impedance measuring instrument.

[0080] Figure 6 This is a graph showing the change in impedance of each channel of the array chip for integrated impedance measurement and microscopic observation as a function of cell line concentration, provided in this embodiment of the present invention.

[0081] like Figure 6 As shown in the figure, the experiment demonstrates that using this array chip can verify that the impedance values ​​of cell lines at different concentrations increase with increasing concentration. Under a microscope, it can also be observed that the number of cells within the wells increases with increasing concentration.

[0082] The integrated impedance measurement and microscopic observation array chip provided in this embodiment can provide a 3D culture environment for cell and organoid cultures, and use cell impedance detection technology curves to characterize cell growth, achieving real-time, label-free, and non-destructive cell monitoring. In addition, it can be placed under a microscope to observe cell growth.

[0083] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this utility model, and not to limit it. Although this utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this utility model.

Claims

1. An array chip integrating impedance measurement and microscopic observation, characterized in that, include: A transparent substrate with a conductive film on one side; A circuit board is disposed on the conductive film, and the circuit board has an array of multiple microchannels. A 3D printing plate is disposed on the side of the circuit board away from the conductive film. The 3D printing plate has an array of multiple microcavities, and the positions of the microcavities correspond one-to-one with the positions of the microchannels. Electrode unit, including: A first electrode plate is placed on the 3D printed plate. The first electrode plate is in contact with the sample in the microchannel. The first electrode plate is adapted to be connected to an impedance measuring instrument for applying an excitation signal to perform impedance detection. A second electrode plate is disposed around the circuit board, and the second electrode plate forms a closed circuit with the first electrode plate through the conductive film.

2. The array chip for integrated impedance measurement and microscopic observation according to claim 1, characterized in that, The transparent substrate and the circuit board are bonded and fixed together by a first UV adhesive layer; And / or, the circuit board and the 3D printed board are bonded and fixed together by a second UV adhesive layer.

3. The array chip for integrated impedance measurement and microscopic observation according to claim 1, characterized in that, The transparent substrate includes either sodium-calcium glass or borosilicate glass.

4. The array chip for integrated impedance measurement and microscopic observation according to claim 1, characterized in that, The conductive film includes an indium tin oxide film.

5. The array chip for integrated impedance measurement and microscopic observation according to claim 1, characterized in that, The circuit board includes a flame-retardant epoxy resin glass fiber laminate.

6. The array chip for integrated impedance measurement and microscopic observation according to claim 1, characterized in that, The 3D printed plate includes a biocompatible resin plate.

7. The array chip for integrated impedance measurement and microscopic observation according to claim 1, characterized in that, The first electrode plate is provided with an internal pin and a first pin; the position of the internal pin corresponds one-to-one with the position of the microcavity, the first pin is symmetrically arranged on both sides of the first electrode plate, and the first pin is connected to the internal pin one-to-one, and the first pin is suitable for connection with an impedance measuring instrument. The second electrode is connected to the conductive film of the transparent substrate by UV adhesive curing, and the second electrode has a second lead of the conductive film.

8. The array chip for integrated impedance measurement and microscopic observation according to claim 7, characterized in that, At least two limiting members are connected to the first electrode by UV adhesive curing, and each pair of limiting members is located at two opposite apex corners of the first electrode. When the first electrode plate is placed on the 3D printing plate, the limiting member abuts against the two opposite sidewalls of the 3D printing plate.

9. The array chip for integrated impedance measurement and microscopic observation according to any one of claims 1 to 8, characterized in that, The microchannel has an aperture of 0.4-1.0 mm, and the center-to-center distance between two adjacent microchannels is 9 mm.

10. The array chip for integrated impedance measurement and microscopic observation according to any one of claims 1 to 8, characterized in that, The diameter of the microcavity is 4-6 mm, the height of the microcavity is 5.4 mm, and the center-to-center distance between two adjacent microcavities is 9 mm.

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