A microfluidic chip for cell function detection

Abstract

The present invention discloses a microfluidic chip for cell function testing, comprising an upper substrate, a middle substrate, and a lower substrate, each provided with a culture medium addition hole and a culture medium discharge hole. Each first flow channel is serially connected to a culture chamber, each culture chamber also correspondingly connected to a second flow channel. A partition is provided in the middle of the culture chamber corresponding to the middle substrate, and the partition is provided with a through hole. A three-layer structure is formed by stacking, and a partition is provided in the culture chamber to separate the culture medium and cell fluid. The cell fluid below the culture chamber partition and the culture medium above the culture chamber partition prevent the cells from flowing out together with the culture medium. Multiple culture chambers, culture medium discharge holes, cell fluid addition holes, and cell fluid discharge holes are provided, facilitating simultaneous drug testing of different cells.

Description

Technical Field

[0001] The invention belongs to the technical field of microfluidic chips, and in particular relates to a microfluidic chip for cell function detection. Background Art

[0002] Microfluidic chip technology, also known as microfluidic chip laboratory or chip laboratory, refers to a chemical or biological laboratory built on a chip of a few square centimeters. It integrates basic operating units involved in fields such as sample preparation, reaction, separation, detection, cell culture, sorting, and lysis onto a very small chip. A network of microchannels is formed, and controllable fluids run through the entire system to achieve biological, chemical, medical diagnosis, and pharmaceutical research and development (this product was originally designed for anti-cancer drug sensitivity and toxicity experiments).

[0003] The basic characteristics and greatest advantages of microfluidic chip technology are: multiple unit structures can be flexibly combined on a tiny chip platform, making the chip design flexible, versatile and fully functional; the tiny internal structural units of the chip require very small amounts of test samples, and the large specific surface area of ​​the microstructure units allows internal reagents to diffuse quickly to achieve rapid reaction and detection; microfluidic chip technology can reduce the technical requirements for medical testing personnel, reduce human errors in testing, and thus reduce the cost of medical testing for patients; because it is completed through instrument automation, more accurate and sensitive test data can be obtained.

[0004] Existing medicine has many kinds of drugs, and in the field of cancer treatment, there is a "cocktail therapy", which specifically tests the anti-cancer effects of different drugs on cancer. Different cancer cells have different stress responses to the same anti-cancer drug. In order to quickly detect the therapeutic effects of different cancer cells on the same drug, it is very unrealistic to conduct human trials. Existing microfluidic chips do not correspond to the same drug to achieve the stress response of different cells, resulting in cumbersome operations. The drug concentration causes errors in the preparation of parallel experiments, resulting in errors in the experimental results. Existing multi-channel microfluidic chips also have the problem of cells mistakenly entering the culture medium flow channel and causing blockages during actual operation. Therefore, the main problem solved by the present invention is that there is currently no microfluidic chip that can perform equivalent detection of the same drug on different cells at the same time. Summary of the Invention

[0005] The purpose of the present invention is to provide a microfluidic chip for cell function detection, which mainly solves the problem that there is currently no microfluidic chip that can perform equivalent detection of the same drug on different cells at the same time.

[0006] To solve the above technical problems, the present invention is achieved through the following technical solutions:

[0007] A microfluidic chip for cell function detection

[0008] The upper substrate, the middle substrate and the lower substrate are sequentially overlapped, abutted and assembled by plasma discharge, and the overall shape is rectangular;

[0009] The upper substrate is provided with a culture medium addition hole and a culture medium outflow hole on opposite sides of the rectangle, and the culture medium outflow holes are multiple and evenly arranged in a row. The culture medium addition hole and the culture medium outflow hole both pass through the upper substrate and extend to the middle substrate. A first flow channel is provided between the culture medium addition hole and the multiple culture medium outflow holes, and the first flow channel is provided on the side of the upper substrate corresponding to the middle substrate;

[0010] The first flow channels are all connected in series with culture cavities, the culture cavities are arranged in a row and correspond one to one with the culture medium outflow holes, the culture cavities penetrate the middle substrate and extend to the upper substrate and the lower substrate respectively;

[0011] Each of the culture chambers is further connected to a second flow channel, the culture chamber is located in the middle of the corresponding second flow channel, and a cell fluid addition hole and a cell fluid discharge hole are respectively provided at both ends of the second flow channel. The cell fluid addition hole and the cell fluid discharge hole both pass through the upper substrate and the middle substrate and extend to the lower substrate, and the second flow channel is provided on a side of the lower substrate corresponding to the middle substrate;

[0012] A partition is provided in the middle of the middle substrate corresponding to the culture cavity, and the partition is provided with a through hole.

[0013] Furthermore, a microfluidic chip for cell function detection is provided, wherein the culture chamber is prismatic and its cross section is rhombus-shaped.

[0014] Furthermore, in a microfluidic chip for cell function detection, the second flow channel is in a herringbone shape, and the cell fluid addition holes and the cell fluid discharge holes are both arranged in a row on both sides of the herringbone shape.

[0015] Furthermore, a microfluidic chip for cell function detection is provided with 8 culture chambers.

[0016] Furthermore, a microfluidic chip for cell function detection is provided, wherein the upper substrate, the middle substrate and the lower substrate are all made of PMMA or PDMS.

[0017] The present invention has the following beneficial effects:

[0018] (1) Different from the previous double-layer structure, a three-layer structure is used here, and a partition is set in the culture chamber to separate the culture medium and the cell fluid. The cell fluid below the culture chamber partition and the culture medium above the culture chamber partition cannot pass through the through hole into the culture medium above, which has the disadvantage of preventing the cells from flowing out together with the culture medium.

[0019] (2) Separating the culture medium and cell fluid is beneficial to the uniformity of the cell fluid. When the cell fluid enters the lower layer of the culture chamber, it is first allowed to stand still so that the cell fluid can solidify under the action of the gel to form a stable spatial structure. The cell fluid is separated by a partition, which is beneficial to the full and uniform contact between the culture medium and the cell fluid, thereby improving the efficiency of cell metabolism.

[0020] (3) Here, there are multiple culture chambers, culture medium outflow holes, cell fluid addition holes, and cell fluid discharge holes, which facilitates simultaneous drug testing of different cells. This is mainly used to test the stress response of cancer cells of different genotypes to the same drug.

[0021] (4) Different from the traditional two-dimensional cell function detection experiment, this chip can keep cells in a non-adherent three-dimensional culture state, and its cell morphology is closer to the state in the human body.

[0022] Of course, any product implementing the present invention does not necessarily need to achieve all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0024] Figure 1 : Structural disassembly diagram of the present invention.

[0025] Figure 2 : Bottom view of the structural diagram of the present invention.

[0026] Figure 3 : An enlarged view of the local structure of the culture chamber of the present invention.

[0027] Figure 4 : Structural diagram of the present invention.

[0028] Figure 5 : Structural diagram of the present invention.

[0029] In the accompanying drawings, the components represented by the reference numerals are as follows:

[0030] Upper substrate 1, middle substrate 2, lower substrate 3, culture medium addition hole 11, culture medium outflow hole 12, first flow channel 13, culture chamber 4, second flow channel 31, cell fluid addition hole 32, cell fluid outflow hole 33, partition 41, through hole 42. DETAILED DESCRIPTION

[0031] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts shall fall within the scope of protection of the present invention.

[0032] In the description of the present invention, it should be understood that the terms "opening", "upper", "lower", "thickness", "top", "middle", "length", "inside", "around" and the like indicating orientation or positional relationship are only for the convenience of describing the present invention and simplifying the description, and do not mean that the components or elements referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention.

[0033] like Figure 1-5 Shown: A microfluidic chip for cell function detection

[0034] The upper substrate 1, the middle substrate 2 and the lower substrate 3 are sequentially overlapped, abutted and combined by plasma discharge, and the overall shape is rectangular;

[0035] The upper substrate 1 is provided with a medium addition hole 11 and a medium outflow hole 12 on opposite sides of the rectangle. The medium is a liquid medium. There are multiple medium outflow holes 12, evenly arranged in rows, while there is a single, larger medium addition hole. Both the medium addition hole 11 and the medium outflow hole 12 pass through the upper substrate 1 and extend to the middle substrate 2. A first flow channel 13 is provided between the medium addition hole 11 and the multiple medium outflow holes 12. The first flow channel 13 is provided on the side of the upper substrate 1 corresponding to the middle substrate 2.

[0036] The first flow channels 13 are all connected in series with culture cavities 4, which are arranged in rows and correspond one-to-one to the culture medium outflow holes 12. The culture cavities 4 pass through the middle substrate 2 and extend to the upper substrate 1 and the lower substrate 3 respectively; here, the culture cavities and the culture medium outflow holes are arranged in rows, which is convenient for marking with a marker.

[0037] Each of the culture chambers 4 is further connected to a second flow channel 31. The culture chamber 4 is located in the middle of the second flow channel 31. A cell fluid addition hole 32 and a cell fluid discharge hole 33 are respectively provided at both ends of the second flow channel 31. The cell fluid addition hole 32 and the cell fluid discharge hole 33 both pass through the upper substrate 1 and the middle substrate 2 and extend to the lower substrate 3. The second flow channel 31 is provided on the side of the lower substrate 3 corresponding to the middle substrate 2.

[0038] The first and second flow channels are circular holes. A template is etched on a polished silicon wafer using a photolithography machine. The template is then cast onto the wafer using the same gel used to create the substrate, creating a biochip with pathways. All the culture medium outflow holes are designed with an etched diameter of 2.5mm. The first flow channel has a diameter of 2.4mm, the cell fluid addition and discharge holes are designed with etched diameters of 2mm, and the second flow channel has a diameter of 1.12mm.

[0039] The middle substrate 2 is provided with a partition 41 in the middle portion corresponding to the culture chamber 4, and the partition 41 is provided with a through hole 42. The designed etching diameter of the middle culture chamber is 1 mm, and the punching diameter of the through hole is 0.1 mm.

[0040] The present invention is a disposable cell culture microfluidic chip structure. Unlike the previous double-layer structure, a three-layer structure is superimposed here. Although the first flow channel and the second flow channel are both connected to the culture chamber, the connection positions are different, and a partition is set in the culture chamber to separate the culture medium and the cell fluid. The cell fluid below the culture chamber partition and the culture medium above the culture chamber partition cannot pass through the through hole into the upper culture medium, which has the disadvantage of preventing the cells from flowing out together with the culture medium.

[0041] Separating the culture medium and cell fluid promotes uniformity of the cell fluid. This is because the cell fluid is injected first, using a pipette to pressurize the fluid. The cell fluid is mixed with gel. When the cell fluid enters the lower layer of the culture chamber, it is allowed to settle to allow the gel to solidify, forming a stable spatial structure. Using a partition to separate the cell fluid prevents it from clogging the primary flow channel and thus hindering the flow of the culture medium. This also facilitates full and uniform contact between the culture medium and the cell fluid, improving cell metabolic efficiency.

[0042] It should be noted that there are multiple culture chambers, culture medium outflow holes, cell fluid addition holes, and cell fluid discharge holes, which facilitate simultaneous drug testing of different cells. This is mainly used for the stress response of different types of cancer cells to the same drug. The drug is dissolved in the culture medium, and a syringe pump is used to continuously supply the culture medium during use. The culture medium that has undergone cell metabolism flows out through the culture medium outflow hole for collection and testing.

[0043] like Figure 3As shown, the culture chamber 4 is prismatic, and its cross section is rhombic. The prismatic culture chamber is advantageous in that there are no bubbles in the center of the cell chamber, thus ensuring that the cells can fully contact the culture medium.

[0044] like Figure 1 As shown, the second flow channel 31 is in a herringbone shape, with the cell fluid inlet 32 ​​and cell fluid outlet 33 arranged in a row on either side of the herringbone. Because the flow channels and perforations of the upper, middle, and lower substrates are all cast, the assembled chip cavity is filled with air. The cell fluid outlet holes are provided for air pressure balance, facilitating smooth and efficient entry of cell fluid into the culture chamber.

[0045] like Figure 1 As shown, the culture chambers 4 are provided with 8 chambers, which are used to perform drug detection on multiple types of cells at the same time.

[0046] The upper substrate 1 , the middle substrate 2 and the lower substrate 3 are all made of PMMA or PDMS.

[0047] It should be noted that PDMS is a biological material, mainly used as a gel for modeling and pouring after photolithography of silicon photoplates. It has a gas exchange function and is the main material used in most cases of this invention. PMMA is a hard material and is not suitable for modeling and pouring after photolithography of silicon photoplates. It has no gas exchange function and requires an additional gas exchange device. The flow channels and perforations set on it are also mechanically engraved, with slight differences in precision, which does not affect the overall effect.

[0048] Throughout this specification, references to terms such as "one embodiment," "example," or "specific example" indicate that the specific features, structures, materials, or characteristics described in conjunction with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, schematic representations of these 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 any one or more embodiments or examples.

[0049] The preferred embodiments of the present invention disclosed above are intended only to help illustrate the present invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the present invention to the specific embodiments described. Obviously, many modifications and variations are possible based on the content of this specification. These embodiments are selected and described in detail in this specification to better explain the principles and practical applications of the present invention, thereby enabling those skilled in the art to better understand and utilize the present invention. The present invention is limited only by the claims and their full scope and equivalents.

[0050] The above embodiments are merely illustrations of the products of the present invention and do not constitute any form of limitation to the present invention. Without departing from the scope of the technical solution of the present invention, any equivalent solution that is simply modified by utilizing the above-disclosed technical contents shall fall within the scope of protection of the present invention.

Claims

1. A microfluidic chip for cell function detection, characterized in that: The invention comprises an upper substrate (1), a middle substrate (2) and a lower substrate (3), wherein the upper substrate (1), the middle substrate (2) and the lower substrate (3) are sequentially overlapped, abutted and assembled by plasma discharge, and the overall shape is rectangular; The upper substrate (1) is provided with a culture medium addition hole (11) and a culture medium outflow hole (12) on opposite sides of the rectangle, the culture medium addition hole (11) is single, and the culture medium outflow hole (12) is multiple and evenly arranged in a row, the culture medium addition hole (11) and the culture medium outflow hole (12) both pass through the upper substrate (1) and extend to the middle substrate (2), a first flow channel (13) is provided between the culture medium addition hole (11) and the multiple culture medium outflow holes (12), and the first flow channel (13) is provided on the side of the upper substrate (1) corresponding to the middle substrate (2); The first flow channels (13) are all connected in series with culture chambers (4), the culture chambers (4) are arranged in a row and correspond one-to-one with the culture medium outflow holes (12), and the culture chambers (4) penetrate the middle substrate (2) and extend to the upper substrate (1) and the lower substrate (3) respectively; The culture chambers (4) are also connected to a corresponding second flow channel (31). The culture chambers (4) are located in the middle of the corresponding second flow channel (31). A cell fluid addition hole (32) and a cell fluid discharge hole (33) are respectively provided at both ends of the second flow channel (31). The cell fluid addition hole (32) and the cell fluid discharge hole (33) are respectively passed through the upper substrate (1) and the middle substrate (2) and extend to the lower substrate (3). The second flow channel (31) is provided on a side of the lower substrate (3) corresponding to the middle substrate (2). The middle portion of the middle substrate (2) corresponding to the culture chamber (4) is provided with a partition (41), and the partition (41) is provided with a through hole (42), and the punching diameter of the through hole is 0.1 mm.

2. The microfluidic chip for cell function detection according to claim 1, characterized in that: The culture chamber (4) is prism-shaped, and its cross section is rhombus-shaped.

3. The microfluidic chip for cell function detection according to claim 1, characterized in that: The second flow channel (31) is in a herringbone shape, and the cell fluid addition holes (32) and the cell fluid discharge holes (33) are both arranged in a row on both sides of the herringbone shape.

4. The microfluidic chip for cell function detection according to claim 1, characterized in that: There are 8 culture chambers (4).

5. The microfluidic chip for cell function detection according to claim 1, characterized in that: The upper substrate (1), the middle substrate (2) and the lower substrate (3) are all made of PMMA or PDMS.

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