A microfluidic device for efficient cell culture and analysis

Abstract

The utility model provides a microfluidic device for efficient cell culture and analysis, which is used to control the flow of fluids, the culture of cells, and the monitoring and control of the cell culture environment; it includes a base plate, a cover plate, a microfluidic chip, and a micropump. The utility model solves the problems of existing microfluidic devices by simplifying the design, improving the accuracy of fluid control, enhancing the ability to simulate the cell microenvironment, integrating real-time monitoring sensors, and optimizing cost-effectiveness. The microfluidic device of the present invention can not only provide a more precise and controllable cell culture environment, but also realize the automation and high-throughput of the cell culture process, thereby promoting the development of cell culture technology. It provides an innovative, efficient, and economical solution for the field of cell culture. The utility model takes into account the functions of cell culture and monitoring, and the core functions include controlling the flow of fluids, the culture of cells, and the monitoring and control of the cell culture environment.

Description

Technical Field

[0001] The utility model relates to the technical field of cell culture devices, in particular to a microfluidic device for efficient cell culture and analysis. Background Art

[0002] Cell culture is a basic technology in the fields of biology, medical research and biopharmaceuticals. It allows scientists to simulate the growth environment of cells in vivo in vitro, conduct research on cell biological characteristics, drug screening, toxicity testing, etc. With the deepening of biomedical research, the requirements for cell culture technology are becoming higher and higher, especially in simulating cell microenvironment and realizing the interaction between cells and cells and cells and extracellular matrix (ECM).

[0003] Traditional cell culture methods, such as using culture dishes, culture flasks or multi-well plates, although successful in some aspects, also have some obvious limitations. First, these methods make it difficult to accurately control the microenvironment in which cells grow, such as oxygen concentration, nutrient supply and removal of metabolic waste. Second, traditional methods usually lack dynamic simulation of the cell microenvironment and cannot simulate the complex and dynamic changes that cells experience in vivo. In addition, these methods often require more manpower and material resources during operation, are costly, and are difficult to achieve high-throughput screening.

[0004] With the continuous deepening of biomedical research, the demand for cell culture technology is also growing. As a cutting-edge technology, microfluidic technology has broad application prospects in the field of cell culture. Future development trends include further improving the integration of microfluidic devices, realizing more complex cell culture models, developing multifunctional integrated microfluidic platforms, and exploring the application of personalized medicine and precision medicine. Microfluidic technology, also known as "Lab-on-a-Chip" technology, is a technology that precisely controls fluid flow at a microscale. The application of this technology in the field of cell culture provides new possibilities for solving the limitations of traditional methods. Microfluidic devices can provide precise fluid control, simulate the microenvironment of cells in the body, realize the interaction between cells and extracellular matrix, and perform high-throughput drug screening and toxicity testing.

[0005] Although microfluidics has shown great potential in the field of cell culture, existing microfluidic devices still have some shortcomings. For example, some devices are complex in design and difficult to manufacture and operate; some devices do not have precise fluid control to meet the strict requirements of cell culture; and some devices lack the ability to monitor and dynamically adjust the cell growth environment in real time. In addition, existing microfluidic devices also face challenges in cost-effectiveness and large-scale production. Utility Model Content

[0006] The utility model aims to provide a microfluidic device for efficient cell culture and analysis with simple structure, high fluid control accuracy, enhanced cell microenvironment simulation capability, integrated real-time monitoring sensors and optimized cost-effectiveness.

[0007] In order to achieve the above purpose, the utility model adopts the following technical solutions:

[0008] A microfluidic device for efficient cell culture and analysis, used for controlling the flow of fluid, cell culture, cell culture environment monitoring and control; comprising a base plate, a cover plate, a microfluidic chip and a micropump; the base plate is provided with a microfluidic chip, the cover plate and the base plate are bonded by a substrate to form a closed microfluidic chip cavity; the microfluidic chip cavity comprises an inlet, a first channel, a first microchamber, a second channel, a second microchamber, a third channel and an outlet connected in sequence; the inlet is used for introducing fluid; the first channel, the second channel and the third channel are used for the path along which the fluid flows; the first microchamber is used for cell culture and analysis; the second microchamber is provided with an integrated sensor for real-time monitoring of the cell growth state and the chemical composition changes in the microenvironment; the outlet is used for collecting cultured cells or waste liquid; the micropump is connected to the outlet through a catheter, and the micropump controls the flow rate and flow direction of the fluid through a microvalve.

[0009] Preferably, one end of the first microchamber is connected to the first channel via a micron flow channel, and the other end of the first microchamber is connected to the second channel via a micron flow channel.

[0010] Preferably, a fourth channel is provided between the first channel and the second channel, and the fourth channel is communicated with the flow channels at both ends of the first microchamber.

[0011] Preferably, a plurality of square microgrooves arranged in a matrix are provided at the bottom of the microchamber cavity of the second microchamber, and the integrated sensor is arranged in the square microgrooves.

[0012] Preferably, the integrated sensor is a colorimetric chemical sensor, comprising at least one colorimetric pigment encapsulated in a silicon-based microsphere.

[0013] Preferably, the base plate and the cover plate are both made of polydimethylsiloxane material, and the cover plate is bonded to the base plate substrate by plasma treatment.

[0014] Compared with the prior art, the beneficial effects of the utility model are as follows:

[0015] (1) The utility model realizes the purpose of cell culture and cell culture environment monitoring by providing a closed microfluidic chip cavity.

[0016] (2) The utility model connects a micro pump at the outlet, and the micro pump can control the flow rate and flow direction of the fluid through a micro valve, thereby achieving convenient operation and high fluid precision control.

[0017] (3) The utility model carves out a microfluidic chip on a plastic material and uses it as a template to cast polydimethylsiloxane, so that the microfluidic chip and the base plate are integrally formed, which has a simple structure and is safely sealed.

[0018] In summary, the utility model solves the problems of existing microfluidic devices by simplifying the design, improving fluid control accuracy, enhancing the ability to simulate the cell microenvironment, integrating real-time monitoring sensors, and optimizing cost-effectiveness. The microfluidic device of the present invention can not only provide a more precise and controllable cell culture environment, but also realize the automation and high-throughput of the cell culture process, thereby promoting the development of cell culture technology. It provides an innovative, efficient and economical solution for the field of cell culture. The utility model takes into account the functions of cell culture and monitoring, and its core functions include controlling the flow of fluids, cell culture, cell culture environment monitoring and control, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 A schematic diagram of a microfluidic device for efficient cell culture and analysis provided by the utility model in application;

[0020] Figure 2 A schematic diagram of the three-dimensional structure of a microfluidic device for efficient cell culture and analysis provided by the utility model;

[0021] Figure 3 The present invention is a schematic diagram of the structure of a microfluidic chip in a microfluidic device for efficient cell culture and analysis provided by the present invention.

[0022] The serial numbers in the figure are as follows:

[0023] 1. Inlet; 2. First channel; 3. First microchamber; 4. Second channel; 5. Second microchamber; 6. Third channel; 7. Outlet; 8. Bottom plate; 9. Cover plate; 10. Injection pump; 11. Micropump; 12. Fourth channel. DETAILED DESCRIPTION

[0024] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments.

[0025] like Figure 2 and Figure 3As shown, the utility model discloses a microfluidic device for efficient cell culture and analysis, which is used to control the flow of fluid, cell culture, cell culture environment monitoring and control, and includes a base plate 8, a cover plate 9, a microfluidic chip and a micropump 10.

[0026] The bottom plate 8 and the cover plate 9 are both made of polydimethylsiloxane material, and the cover plate 9 is bonded to the bottom plate 8 substrate by plasma treatment to form a closed microfluidic chip cavity.

[0027] The microfluidic chip cavity comprises an inlet 1, a first channel 2, a first microchamber 3, a second channel 4, a second microchamber 5, a third channel 6 and an outlet 7 which are connected in sequence. Further, in this embodiment, a fourth channel 12 is provided between the first channel 2 and the second channel 4, and the fourth channel 12 communicates with the flow channels at both ends of the first microchamber 3.

[0028] The inlet 1 introduces fluid through the syringe pump 11. The first channel 2, the second channel 4, the third channel 6 and the fourth channel 12 are used for the path along which the fluid flows; the first microchamber 3 is used for cell culture and analysis; the second microchamber 5 is provided with an integrated sensor for real-time monitoring of the cell growth state and the chemical composition changes in the microenvironment; the outlet 7 is used to collect the cultured cells or waste liquid; the micropump 10 is connected to the outlet 7 through a catheter, and the micropump 10 controls the flow rate of the fluid and selects the flow direction into different channels through two sets of microvalves.

[0029] Further, in the present embodiment, the first microchamber 3 is smaller than the second microchamber 5, the first microchamber adopts a direct 2000 micron cavity, and the second microchamber 5 adopts a direct 5000 micron cavity. One end of the first microchamber 3 is connected to the first channel 2 through a 500 micron flow channel, and the other end of the first microchamber 3 is connected to the second channel 4 through a 150 micron flow channel. The channel structure of the first microchamber 3, which is wide in front and narrow in the back, can ensure that after the cells form a large three-dimensional cell ball or organoid structure, they can stay in the first microchamber 3 and are not easily flowed away by the movement of the fluid. Due to the above-mentioned flow-limiting structure, the fluid that cannot flow quickly can be connected to the second channel 4 through the fourth channel 12, thereby realizing the flow of the fluid.

[0030] The bottom of the microchamber cavity of the second microchamber 5 is provided with a plurality of square microgrooves arranged in a matrix, and the integrated sensors are arranged in the square microgrooves.

[0031] Furthermore, the integrated sensor in this embodiment adopts a colorimetric chemical sensor, including encapsulating one or more colorimetric pigments (cresol red, phenol red) in silicon-based microspheres, and then loading the silicon-based microspheres encapsulating the colorimetric pigments into the square microgrooves at the bottom of the integrated sensor chamber, and observing them through an optical detection instrument. In this embodiment, the optical detection instrument adopts a spectrometer, and an external spectrometer detects color changes during detection, which is used to detect parameters such as carbon dioxide concentration and pH value in the microenvironment.

[0032] like Figure 1 As shown, the preparation process of the utility model is as follows:

[0033] Step S1: Carve the shape template of the microfluidic chip channel and microchamber on the PMMA material by laser direct writing process.

[0034] Step S2: Cast PDMS material on the PMMA template to form a bottom plate 8. After solidification, the bottom plate 8 with complete channels and microchambers of the microfluidic chip is obtained. Then, a cover plate 9 made of PDMS material is taken, and after the surface is plasma treated, it is bonded to the bottom plate 8 substrate containing channels and microchambers to form a closed microfluidic chip cavity.

[0035] Step S3: When needed, holes are drilled at the inlet 1 and the outlet 7, and the culture medium is injected into the inlet 1. The integrated sensor of the second microchamber 5 is connected to an external optical detector for analysis and observation, and the outlet 7 is connected to a micropump, which is used to accurately control the fluid flow and flow rate in the microfluidic chip cavity through a microvalve.

[0036] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0037] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. In the description of the present utility model, the meaning of "plurality" is two or more, unless otherwise clearly and specifically defined.

[0038] The above are only preferred specific implementation methods of the utility model, but the protection scope of the utility model is not limited to this. Any technician familiar with the technical field within the technical scope disclosed by the utility model, who makes equivalent replacements or changes based on the technical scheme and utility model concept of the utility model, should be covered by the protection scope of the utility model.

Claims

1. A microfluidic device for efficient cell culture and analysis, used for controlling fluid flow, cell culture, cell culture environment monitoring and control; characterized in that: It comprises a base plate (8), a cover plate (9), a microfluidic chip and a micro pump (10); The bottom plate (8) is provided with a microfluidic chip, and the cover plate (9) is bonded to the bottom plate (8) to form a closed microfluidic chip cavity; The microfluidic chip cavity comprises an inlet (1), a first channel (2), a first microchamber (3), a second channel (4), a second microchamber (5), a third channel (6) and an outlet (7) which are connected in sequence; The inlet (1) is used to introduce fluid; the first channel (2), the second channel (4) and the third channel (6) are used for the path along which the fluid flows; the first microchamber (3) is used for cell culture and analysis; the second microchamber (5) is provided with an integrated sensor, and an external optical detector is used to monitor the cell growth state and the chemical composition changes in the microenvironment in real time; the outlet (7) is used to collect cultured cells or waste liquid; The micro pump (10) is connected to the outlet (7) via a conduit, and the micro pump (10) controls the flow rate and flow direction of the fluid via a micro valve.

2. A microfluidic device for efficient cell culture and analysis according to claim 1, characterized in that: One end of the first microchamber (3) is connected to the first channel (2) via a 500-micrometer flow channel, and the other end of the first microchamber (3) is connected to the second channel (4) via a 150-micrometer flow channel.

3. A microfluidic device for efficient cell culture and analysis according to claim 2, characterized in that: A fourth channel (12) is provided between the first channel (2) and the second channel (4), and the fourth channel (12) is communicated with the flow channels at both ends of the first microchamber (3).

4. A microfluidic device for efficient cell culture and analysis according to claim 1, characterized in that: The bottom of the microchamber cavity of the second microchamber (5) is provided with a plurality of square microgrooves arranged in a matrix, and the integrated sensor is arranged in the square microgrooves.

5. A microfluidic device for efficient cell culture and analysis according to claim 4, characterized in that: The integrated sensor adopts a colorimetric chemical sensor, which includes encapsulating at least one colorimetric pigment in a silicon-based microsphere.

6. The microfluidic device for efficient cell culture and analysis according to claim 1, characterized in that: The base plate (8) and the cover plate (9) are both made of polydimethylsiloxane material, and the cover plate (9) is bonded to the base plate (8) substrate by plasma treatment.

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