Automatic microfluid cell culture system and method integrated with artificial intelligence

Abstract

The invention relates to an automatic microfluid cell culture system and method integrated with artificial intelligence, and belongs to the field of biomedical engineering. Through a modular micro-fluidic chip and an advanced monitoring algorithm, efficient and automatic management of a cell culture process is realized. The system analyzes cell behaviors in real time by utilizing machine learning, automatically adjusts culture conditions, and ensures consistency and stability of cell growth. The application of the digital microfluidic technology allows accurate control of single cells or microspheres, and optimizes the high-throughput screening process. In addition, the system supports 3D cell culture and organoid model construction, and provides an experimental platform closer to physiological conditions. Through integration of automatic operation and remote monitoring functions, cell culture is more convenient and controllable, and the efficiency and accuracy of biomedical research and drug screening are remarkably improved. The design of the system pays attention to environmental protection and sustainability, resource consumption is reduced, and innovativeness and practicability in a cell culture technology are shown.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical engineering, and in particular relates to an automated microfluidic cell culture system and method integrated with artificial intelligence. Background Art

[0002] Cell culture is a basic and key technology in biomedical research, and is widely used in many fields such as drug screening, disease mechanism research, tissue engineering, and production of biological products. Traditional cell culture methods usually rely on manual operations, including cell inoculation, culture medium replacement, gas environment control, and cell harvesting. These operations are not only time-consuming and labor-intensive, but also easily affected by human factors, resulting in inconsistent culture conditions and poor reproducibility of results.

[0003] With the development of microfluidic technology, the field of cell culture has begun to show a trend of automation and miniaturization. Microfluidic systems can accurately control fluids on a microscale, providing cells with a controllable and stable culture environment. However, existing microfluidic cell culture systems are often complex in structure, expensive, and lack sufficient flexibility and intelligence in operation, which limits their widespread adoption in scientific research and clinical applications.

[0004] In order to overcome these limitations, researchers have begun to explore the combination of artificial intelligence technology and microfluidic systems in recent years to achieve automated monitoring and intelligent management of the cell culture process. Artificial intelligence algorithms, especially machine learning and image recognition technology, can be used to analyze the morphology and growth state of cells and automatically adjust culture conditions to improve the efficiency and quality of cell culture. In addition, the application of digital microfluidics technology makes it possible to precisely manipulate single cells or cell groups, providing a new approach for single-cell analysis and high-throughput screening.

[0005] Although these technological advances have brought new opportunities to the field of cell culture, there is still the challenge of how to integrate these highly specialized technologies into a user-friendly, cost-effective system. Therefore, the development of a new type of automated microfluidic cell culture system with integrated artificial intelligence, which can not only provide precise cell culture environment control, but also realize remote monitoring and intelligent management, has become an important research direction in this field. It is in this context that the present invention came into being, aiming to promote the development of cell culture technology and meet the needs of modern biomedical research through innovative system design and intelligent algorithms. Summary of the invention

[0006] The purpose of the present invention is to provide an automated microfluidic cell culture system and method integrated with artificial intelligence, which can realize automation, precise control and intelligent management of the cell culture process to improve the efficiency, accuracy and repeatability of cell culture. The design of the system allows users to quickly adjust and replace components according to different experimental requirements, while real-time monitoring and optimization of cell culture conditions through artificial intelligence algorithms.

[0007] To achieve the above purpose, the technical solution of the present invention is: an automated microfluidic cell culture system with integrated artificial intelligence, including: a microfluidic chip, an artificial intelligence control unit, an automated operation module, a digital microfluidic module, a remote monitoring platform, a 3D cell culture module, and a self-circulating heat dissipation system; wherein the microfluidic chip is a core component and is closely connected to the artificial intelligence control unit through a standardized interface. The artificial intelligence control unit has an embedded machine learning algorithm, which can analyze the cell culture data from the microfluidic chip in real time, such as cell growth status and metabolite concentration, and automatically adjust the culture conditions, including temperature, pH value and oxygen concentration, to ensure that the cells grow in the best environment. The automated operation module is connected to the microfluidic chip through a precise piping system and is responsible for performing liquid delivery and environmental control operations. The module integrates precision pumps, valves, temperature controllers and gas supply systems, and accurately performs tasks such as liquid replacement, nutrient addition and waste removal during cell culture according to the instructions of the artificial intelligence control unit. Digital microfluidic technology is combined with the microfluidic channel of the microfluidic chip, and non-contact manipulation of cells is achieved by using electrowetting or dielectrophoresis drive mechanisms. The application of this technology improves the accuracy and repeatability of cell manipulation, allowing for the precise capture, separation and analysis of single cells or microspheres. The remote monitoring platform is connected to the artificial intelligence control unit via a wireless or wired network, providing a user interface and data management system. Researchers can use the platform to monitor cell culture parameters and historical data in real time, achieve remote monitoring and operation, and improve the convenience and efficiency of experiments. The 3D cell culture module is combined with a microfluidic chip, using scaffolds and biomaterials to form a three-dimensional cell structure within the chip. This module can simulate a more complex cell microenvironment, provide cells with a growth environment that is closer to in vivo conditions, and enhance the physiological relevance of cell culture. The self-circulating heat dissipation system is connected to the microfluidic chip through a microfluidic heat sink and a temperature sensor to maintain a constant temperature of the chip. The system adjusts the chip temperature through a thermoelectric module or a microfluidic heat sink to ensure the temperature stability of the cell culture process and provide a stable growth environment for cells. Finally, environmentally friendly materials and energy-saving designs run through the entire system, from the manufacture of microfluidic chips to the optimization of energy efficiency, reflecting the sustainability of the system. These designs not only reduce the impact on the environment, but also improve the overall energy efficiency of the system, ensuring the automation, accuracy and environmental stability of the cell culture process. Through the efficient integration and intelligent connection of the above modules, the system of the present invention provides a powerful tool for biomedical research and drug development.

[0008] The microfluidic chip adopts a modular design to achieve rapid adaptation to different cell culture needs. Each chip unit has independent microfluidic channels and reaction chambers. The dimensions of these channels are customized according to the cell type and culture requirements. For example, the channel width may be designed to range from 50 microns to 500 microns to accommodate cells of different sizes. The system has designed a variety of modular chip units, including standard units for basic cell culture, dedicated units for specific cell types, and advanced units for complex cell models, which can be quickly replaced through automated operation modules. The automated operation module is controlled by a precise robotic arm and automated software, and can identify and grab the correct chip unit and install it in the microfluidic chip slot. This process can be completed in a few minutes. Digital microfluidics plays a key role in the system. It precisely manipulates cells and liquids in microfluidic channels through electrowetting or dielectrophoresis drive mechanisms, making the cell culture process more refined and controllable. The remote monitoring platform provides a user-friendly interface, allowing researchers to access and control the cell culture system through the Internet from anywhere in the world, view the cell culture status in real time, adjust the culture parameters, and even replace the chip unit remotely when necessary, greatly improving the convenience and flexibility of the system. Through the efficient integration and intelligent connection of these modules, the system of the present invention ensures the automation, accuracy and environmental stability of the cell culture process, providing a powerful tool for biomedical research and drug development.

[0009] The AI ​​control unit uses integrated image recognition software to perform high-precision analysis of cell morphology and growth status. Using advanced machine learning algorithms, the software is able to identify and quantify multiple morphological features of cells, such as cell area, perimeter, shape factor, and different stages of the cell cycle. By setting thresholds, the software can automatically distinguish healthy cells from damaged cells, as well as actively dividing cells from inactive cells. For example, the software can identify that the average value of cell area is approximately 500 square microns, and evaluate the growth status of cells based on the distribution of cell perimeters.

[0010] The image recognition software receives image data captured by a high-resolution microscope camera embedded in the microfluidic chip. The camera automatically captures images of the cell culture area at a frequency of once every 10 minutes, ensuring continuous monitoring of the cell culture process. The artificial intelligence control unit analyzes these images in real time and transmits the analysis results to the remote monitoring platform for researchers to make further evaluations and decisions. During the cell culture process, the artificial intelligence control unit will also automatically adjust the culture conditions according to changes in cell morphology, such as adjusting the flow and composition of the culture medium to optimize the growth environment of the cells. For example, when the software recognizes that the cell density exceeds a predetermined threshold, it will automatically instruct the automated operation module to increase the frequency of culture medium replacement to avoid overcrowding of cells. The artificial intelligence control unit is also able to predict possible problems in the cell culture process, such as contamination or cell stress response, through image recognition software, and take timely measures to intervene. This intelligent monitoring and management greatly improves the efficiency and success rate of cell culture, providing strong technical support for biomedical research.

[0011] The automated operation module is equipped with an automatic sampler and an analyzer, which are specially used for sample collection and instant analysis during cell culture. The automatic sampler can accurately collect cell samples from the reaction chamber of the microfluidic chip according to the preset program, and the sampling error is controlled within ±10%, ensuring the representativeness of the sample and the accuracy of the analysis. The sampled cell samples are automatically transported to the analyzer, which integrates a variety of sensors and detectors to quickly measure key parameters such as pH, dissolved oxygen (DO), carbon dioxide (CO2) concentration in the cell culture medium, as well as cell density and metabolite concentration. The measurement parameters of the analyzer are set as follows: the measurement range of pH is 6.5 to 8.5, with an accuracy of ±0.05 pH units; the measurement range of dissolved oxygen is 0 to 100% air saturation, with an accuracy of ±1% air saturation; the measurement range of carbon dioxide concentration is 0 to 50%, with an accuracy of ±2%. The measurement of cell density is carried out by capacitance method, with a measurement range of 0 to 5×10^6 cells / mL and an accuracy of ±5%. Real-time monitoring of these parameters is essential for evaluating the growth status of cells and adjusting culture conditions. The integration of automatic samplers and analyzers makes sample collection and analysis in the cell culture process more efficient and automated. Through the automated operation module, researchers can achieve real-time monitoring and rapid response to the cell culture status without interfering with the cell culture environment, thereby optimizing the cell culture process and improving the success rate of cell culture and the reliability of experimental data. This automated and intelligent cell culture system provides strong technical support for biomedical research and drug development.

[0012] In the automated microfluidic cell culture system of the present invention, the digital microfluidic module realizes non-contact manipulation of cells or microspheres by integrating an acoustic wave generator. The acoustic wave generator can generate acoustic waves of specific frequency and amplitude, which form a standing wave field in the liquid medium of the microfluidic chip, and use the radiation pressure of the acoustic wave to accurately manipulate the cells. For example, the acoustic wave generator can be set to generate a frequency of 10 MHz and an amplitude of 0.5 mm to form enough sound pressure to manipulate cells without causing damage to the cells. By adjusting the frequency and amplitude of the acoustic wave generator, operations such as cell aggregation, dispersion, translation and rotation can be achieved. The integration of the acoustic wave generator and the microfluidic chip is achieved by precise alignment and connection to ensure that the acoustic wave energy is effectively transferred to the liquid medium. During the cell culture process, the output power of the acoustic wave generator can be adjusted within the range of 0.1 to 1 watt to meet the manipulation requirements of different cell types. By adjusting the frequency of the acoustic wave generator to 10 MHz and setting the amplitude to 0.5 mm, the aggregation operation of the cell cluster can be achieved. This acoustic wave manipulation technique provides a non-invasive, high-throughput method for cell culture and analysis with broad application prospects.

[0013] The remote monitoring platform uses advanced network technology to achieve real-time monitoring of cell culture status. Users can access the platform anytime and anywhere through mobile devices such as smartphones or tablets. The platform supports encrypted data transmission through secure network protocols such as HTTPS to ensure data security and privacy. Users can view images and data of cell culture in real time on mobile devices through dedicated applications, including key parameters such as cell density, growth rate and chemical composition of culture medium.

[0014] The user interface design of the remote monitoring platform is intuitive and easy to use. Researchers can remotely control the cell culture environment through simple operations, such as adjusting temperature, pH value and gas concentration. The platform also provides an alarm system. When the cell culture conditions exceed the preset range, the system will automatically send a notification to the user to ensure timely measures. For example, the system can set the temperature alarm threshold to 37°C ± 0.5°C and the pH alarm threshold to 7.4 ± 0.1 to ensure the stability of the cell culture environment. The remote monitoring platform can also store and analyze long-term accumulated cell culture data to help researchers optimize experimental plans and improve experimental efficiency. Through big data analysis and machine learning algorithms, the platform can predict cell growth trends and culture effects, providing strong data support for scientific research. This remote monitoring and management capability allows researchers to access and control the cell culture system through the Internet at any location, view the cell culture status in real time, adjust the culture parameters, and even remotely replace the chip unit when necessary, greatly improving the convenience and flexibility of the system. Through the efficient integration and intelligent connection of these modules, the system of the present invention ensures the automation, accuracy and environmental stability of the cell culture process, providing a powerful tool for biomedical research and drug development.

[0015] The 3D cell culture module is capable of constructing complex tissue models to simulate the environment of real biological tissues through its multi-layer structural design. The module uses an innovative multi-layer hydrogel technology, in which the thickness of each layer of hydrogel can be controlled between 100 microns and 500 microns to meet the attachment and growth requirements of different cell types. By precisely controlling the hydrogel composition and porosity of each layer, a microenvironment with specific mechanical properties and biochemical signals can be created to promote cell proliferation and differentiation. In the process of constructing tissue models, the 3D cell culture module can achieve a gradient distribution of oxygen, nutrients, metabolites and soluble signals to form a diverse cell population. For example, by adjusting the oxygen permeability in the hydrogel, the oxygen gradient of different tissues in the body can be simulated, thereby affecting the behavior and function of cells. In addition, the module is also able to simulate cell-to-cell interactions, including three-dimensional cell-to-cell contact and direct or indirect cell-to-cell communication, which is crucial for studying the behavior of cells in the body. In order to achieve these complex 3D structures, the 3D cell culture module uses a combination of multiple cell types and biomaterials. For example, a natural ECM gel derived from the basement membrane of mouse sarcoma cells can be used, the main components of which include laminin, type IV collagen, proteoglycan heparan sulfate and nestin, which can provide rich nutrition and are highly compatible with cell development. By planting cells in these prefabricated hydrogel layers, a tissue model with a specific structure and function can be formed. Through the automated operation module, researchers can easily replace the 3D cell culture module to adapt to different experimental needs. This modular design not only improves the flexibility of the experiment, but also simplifies the experimental operation, making the 3D cell culture technology more efficient and user-friendly. Through these innovative designs and functions, the system of the present invention provides a powerful tool for biomedical research and drug development, especially in the fields of tissue engineering and regenerative medicine, showing great application potential.

[0016] The self-circulating heat dissipation system significantly improves the temperature control capability by integrating thermoelectric modules. Thermoelectric modules, also known as Peltier elements, use the Peltier effect to achieve heat movement. According to the direction of the current, one side absorbs heat and the other side releases heat, thereby achieving precise temperature control. This module can quickly adjust the temperature according to the specific needs of cell culture to ensure the stability of the cell culture environment. The design specifications of the thermoelectric module can be customized to meet different cell culture needs. For example, the size of the module can range from 20mm to 40mm to accommodate culture chambers of different sizes. The thermoelectric module can be built-in with a temperature sensor, such as a thermistor, to achieve high-precision temperature control with an accuracy of ±0.05°C. In addition, a heat sink or heat sink can be connected to the surface of the module to enhance the heat dissipation effect, which is essential for maintaining a constant temperature during cell culture. During the cell culture process, the thermoelectric module can work in conjunction with the heat sink and temperature sensor of the microfluidic chip to monitor and adjust the temperature of the chip in real time through the control of the automated operation module. When the chip temperature exceeds the preset range, the thermoelectric module will automatically start, and by adjusting the direction and size of the current, the chip temperature will be quickly adjusted back to a range suitable for cell growth, such as maintaining a physiological temperature of 37°C ± 0.1°C. The application of thermoelectric modules is not limited to temperature control, but can also be used for thermal management in the cell culture process, such as maintaining a constant temperature environment in a cell culture incubator, or providing necessary thermal cycles during cell culture. This modular design allows the thermoelectric modules to be quickly replaced and adjusted according to different experimental requirements, thereby improving the flexibility and adaptability of the cell culture system. Through the integration of thermoelectric modules, the automated microfluidic cell culture system of the present invention can achieve precise control of the cell culture environment, providing a stable and controllable temperature condition for cell culture, thereby improving the efficiency and quality of cell culture. The design of this self-circulating heat dissipation system not only enhances the temperature control capability, but also provides new possibilities for the development of cell culture technology.

[0017] The system of the present invention also includes environmentally friendly material design and energy-saving design process, and the sustainability of the system is significantly improved by integrating solar panels. These solar panels are designed to match the energy needs of the system, providing renewable energy and reducing dependence on traditional power grids. The peak power output of the solar panels is set at 300W, which is sufficient to meet the energy needs of the system under sufficient sunlight conditions. In the system design, the size and angle of the solar panels are optimized to maximize the absorption of solar energy while taking into account no shadow or reflection interference to the surrounding environment. The solar panels are connected to other components of the system through an efficient energy management system that can intelligently distribute and regulate energy to ensure the continuity and stability of the cell culture process. In insufficient sunlight or at night, the system automatically switches to backup energy sources such as battery storage or grid power supply to ensure uninterrupted cell culture. The integration of solar panels also helps to reduce the carbon footprint of the system, which is in line with the concept of green building and sustainable development. Through this design, the system not only improves energy efficiency, but also demonstrates the possibility of implementing environmental protection and energy-saving measures in biomedical research facilities. This innovative environmentally friendly material and energy-saving design make the automated microfluidic cell culture system a model of environmental protection and energy saving while providing high-precision cell culture.

[0018] The present invention also provides an automated microfluidic cell culture method based on the above-mentioned system with integrated artificial intelligence, comprising the following steps: S1. Initialize the microfluidic chip: First, install the microfluidic chip into the system and sterilize it with ultraviolet radiation for 30 minutes to ensure that the chip surface is sterile. Then, inject the basic culture medium preheated to 37°C into the microfluidic channel of the chip through the precision pump of the automated operation module, and control the flow rate at 0.1 mL / min to ensure that the channel is fully infiltrated and free of bubbles.

[0019] S2. Precise distribution of cells or microspheres: Using digital microfluidics technology, cells or microspheres are precisely distributed to specific areas of the microfluidic chip at a flow rate of 0.05 mL / min through electrowetting or dielectrophoresis drive mechanisms. Approximately 10,000 cells are inoculated in each chamber to ensure uniform cell distribution.

[0020] S3, Automatic culture medium replacement and nutrient addition: Through the automated operation module, the cell culture medium is automatically replaced, nutrients are added, and waste is removed. The culture medium replacement frequency is set to once every 24 hours, and nutrient addition is adjusted according to cell growth needs, usually once every 48 hours.

[0021] S4. Monitor and adjust the culture environment: Use an artificial intelligence control unit to monitor the cell culture environment, including maintaining the temperature at 37°C ± 0.1°C, controlling the pH value at 7.4 ± 0.05, and setting the oxygen concentration to 5% CO2. These parameters are monitored in real time by sensors and automatically adjusted to the set values.

[0022] S5. Remote observation and operation: Through the remote monitoring platform, researchers can observe the cell status in real time, including cell morphology, growth rate and cell density, and perform necessary operations. The platform provides images and data updated every 10 minutes to ensure that researchers can respond in time.

[0023] S6. Construction and cultivation of organoid models: In the 3D cell culture module, scaffolds and biomaterials are used to construct three-dimensional cell structures to simulate complex biological tissue structures. The temperature of the module is maintained at 37°C, the pH value is controlled at 7.2, and the oxygen concentration is 20% O2 to simulate the in vivo environment.

[0024] S7. Maintain constant temperature: Use a self-circulating cooling system, microfluidic heat sink and temperature sensor to maintain a constant temperature of the chip. The thermal conductivity of the heat sink is set to 200 W / mK to ensure rapid heat dissipation.

[0025] S8. Sample collection and immediate analysis: Regularly collect samples and perform immediate analysis during the cell culture process through the automated operation module to evaluate the cell culture effect. The sample collection cycle is once every 72 hours, and the analysis includes cell viability, metabolic activity, and protein expression levels.

[0026] S9. Data analysis and culture program optimization: The artificial intelligence control unit is used to analyze the collected data and optimize the subsequent cell culture program. The analysis results are used to adjust the culture medium formula, nutrient concentration and culture conditions.

[0027] S10. Collecting cultures: After cell culture is completed, collect the cultures, including cell layers, cell spheres, or organoid models, through the automated operation module. During the collection process, ensure aseptic operation to avoid cell contamination.

[0028] S11. Subsequent analysis: The collected cultures are subjected to subsequent biological, pharmacological or toxicological analysis to evaluate their function and safety. The analysis includes gene expression analysis of cells, drug response testing and toxicity assessment to ensure the quality and suitability of the cultures.

[0029] Through the above steps, the automated microfluidic cell culture system of the present invention can provide a standardized, automated cell culture solution, which is applicable to multiple fields such as biomedical research, drug screening, tissue engineering and disease model research.

[0030] Compared with the prior art, the present invention has the following beneficial effects: the design of the system of the present invention focuses on environmental protection and sustainability, reduces resource consumption, and demonstrates innovation and practicality in cell culture technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 A schematic diagram of the automated microfluidic cell culture system of the present invention. Figure 2 The cell culture method flow chart of the present invention. DETAILED DESCRIPTION

[0032] The following will be combined with the attached embodiment of the present invention Figure 1 and Figure 2 , the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] In the automated microfluidic cell culture system of the present invention, the microfluidic chip, as the core of the system, integrates microfluidic channels, reaction chambers, gas exchange areas and waste collection systems for cell culture to provide cells with a suitable growth environment. The width and depth of the microfluidic channel are designed to be 100 microns to accommodate the size and fluid dynamics of the cells. The volume of the reaction chamber is controlled at the nanoliter level to achieve precise chemical and biological reaction control. The gas exchange area exchanges gas with the outside through a microporous membrane to ensure that the oxygen and carbon dioxide levels in the cell culture environment are maintained under physiological conditions of 37°C and 5% CO2. The waste collection system effectively removes metabolic wastes at a flow rate of 0.1 mL / min through a micropump.

[0034] The artificial intelligence control unit is closely connected to the microfluidic chip, and analyzes the cell culture data from the chip through an integrated machine learning algorithm, and automatically adjusts the culture conditions in real time. The unit monitors the culture environment in real time through temperature sensors and pH sensors, and automatically adjusts the temperature controller and pH regulator through a feedback control mechanism to maintain the temperature of the cell culture environment within the range of 37℃±0.1℃ and the pH value within the range of 7.4±0.05. The automated operation module interfaces with the microfluidic chip through precision pumps, valves, temperature controllers, and gas supply systems to perform precise liquid delivery and environmental control operations. The flow rate of the precision pump is adjustable from 0.01 mL / min to 1 mL / min to meet different culture needs. The valve uses high-precision electromagnetic control to ensure accurate switching and addition of culture medium. Digital microfluidics technology is combined with the microfluidic channels of the microfluidic chip to achieve non-contact manipulation of cells using electrowetting or dielectrophoresis drive mechanisms. The electrowetting drive mechanism changes the surface tension of the liquid by applying voltage on both sides of the microfluidic channel, thereby achieving precise movement and manipulation of droplets. The dielectrophoresis drive mechanism uses electric fields to act on charged particles to achieve precise manipulation of cells or microspheres. The remote monitoring platform is connected to the artificial intelligence control unit through a wireless or wired network, providing a user interface and a data management system, allowing researchers to monitor and record cell culture parameters and historical data in real time. The user interface provides an intuitive operation interface, allowing users to set culture parameters, view real-time data and historical trends, and store and analyze data through the data management system. The 3D cell culture module is combined with a microfluidic chip to form a three-dimensional cell structure in the chip using a scaffold and biomaterials. The scaffold material is selected as polylactic-co-glycolic acid (PLGA) with good biocompatibility and a pore size of 20-40 microns to support cell attachment and growth. The self-circulating heat dissipation system is connected to the microfluidic chip through a microfluidic heat sink and a temperature sensor to maintain a constant temperature of the chip. The heat sink is made of metal alloy material with high thermal conductivity to ensure rapid conduction and dissipation of heat. Finally, the environmentally friendly materials and energy-saving design run through the entire system, from the manufacture of microfluidic chips to the optimization of energy efficiency, reflecting the sustainability of the system. The chip material is biodegradable bioplastic, which reduces the impact on the environment. The energy efficiency is optimized by integrating solar panels, reducing the dependence on external power sources, reflecting the green and energy-saving concept of the system.

[0035] Microfluidic chips can quickly adapt to different cell culture needs by adopting a modular design. Each chip unit has independent microfluidic channels and reaction chambers. The dimensions of these channels are customized according to the cell type and culture requirements. For example, the channel width may be designed to range from 50 microns to 500 microns to accommodate cells of different sizes. The system has designed a variety of modular chip units, including standard units for basic cell culture, dedicated units for specific cell types, and advanced units for complex cell models, which can be quickly replaced through automated operation modules. The automated operation module is controlled by a precise robotic arm and automated software, and can identify and grab the correct chip unit and install it in the microfluidic chip slot. This process can be completed in a few minutes. Digital microfluidics plays a key role in the system. It precisely manipulates cells and liquids in microfluidic channels through electrowetting or dielectrophoresis drive mechanisms, making the cell culture process more refined and controllable. The remote monitoring platform provides a user-friendly interface, allowing researchers to access and control the cell culture system through the Internet from anywhere in the world, view the cell culture status in real time, adjust the culture parameters, and even replace the chip unit remotely when necessary, greatly improving the convenience and flexibility of the system. Through the efficient integration and intelligent connection of these modules, the system of the present invention ensures the automation, accuracy and environmental stability of the cell culture process, providing a powerful tool for biomedical research and drug development.

[0036] The AI ​​control unit integrates advanced image recognition software that can analyze cell morphology and growth status. By setting specific algorithm parameters, such as a cell area threshold of 500 square microns and a cell perimeter threshold, the image recognition software can accurately identify and quantify multiple morphological features of cells. For example, the software can identify the average area of ​​cells and evaluate the growth status of cells based on the distribution of cell perimeters. In addition, the software can monitor the physiological activities of cells by analyzing the intensity of the fluorescent marker signal of the cells, and the threshold of the signal intensity may be set within a specific range of fluorescence units. The AI ​​control unit receives real-time image data of the cell culture area by connecting to a high-resolution microscope camera on the microfluidic chip. The camera automatically takes images at preset time intervals, such as every 10 minutes, to ensure continuous monitoring of the cell culture process. The control unit analyzes these images in real time and transmits the analysis results to the remote monitoring platform for researchers to make further evaluations and decisions. In this way, researchers can access and control the cell culture system via the Internet from any location, view the cell culture status in real time, adjust the culture parameters, and even replace the chip unit remotely when necessary. In addition, the AI ​​control unit can also automatically adjust the culture conditions according to changes in cell morphology, such as adjusting the flow and composition of the culture medium. For example, when the software recognizes that the cell density exceeds a predetermined threshold, it will automatically instruct the automated operation module to increase the frequency of culture medium replacement to avoid overcrowding of cells. This intelligent monitoring and management greatly improves the efficiency and success rate of cell culture, providing strong technical support for biomedical research.

[0037] The automated operation module is equipped with an automatic sampler and analyzer, which are specially designed for sample collection and instant analysis during cell culture. The automatic sampler can accurately collect cell samples from the reaction chamber of the microfluidic chip according to the preset program, and the sampling error is controlled within ±10%, ensuring the representativeness of the sample and the accuracy of the analysis. The sampled cell samples are automatically transported to the analyzer, which integrates a variety of sensors and detectors to quickly measure key parameters such as pH, dissolved oxygen (DO), carbon dioxide (CO2) concentration in the cell culture medium, as well as cell density and metabolite concentration. The measurement parameters of the analyzer are set as follows: the measurement range of pH is 6.5 to 8.5, with an accuracy of ±0.05 pH units; the measurement range of dissolved oxygen is 0 to 100% air saturation, with an accuracy of ±1% air saturation; the measurement range of carbon dioxide concentration is 0 to 50%, with an accuracy of ±2%. The measurement of cell density is carried out by capacitance method, with a measurement range of 0 to 5×10^6 cells / mL and an accuracy of ±5%. Real-time monitoring of these parameters is essential for evaluating the growth status of cells and adjusting culture conditions. The integration of automatic samplers and analyzers makes sample collection and analysis in the cell culture process more efficient and automated. Through the automated operation module, researchers can achieve real-time monitoring and rapid response to the cell culture status without interfering with the cell culture environment, thereby optimizing the cell culture process and improving the success rate of cell culture and the reliability of experimental data. This automated and intelligent cell culture system provides strong technical support for biomedical research and drug development.

[0038] The digital microfluidic module achieves non-contact manipulation of cells or microspheres by integrating an acoustic wave generator. The acoustic wave generator can generate acoustic waves of specific frequency and amplitude, which form a standing wave field in the liquid medium of the microfluidic chip, and use the radiation pressure of the acoustic wave to precisely manipulate the cells. For example, the acoustic wave generator can be set to generate a frequency of 10 MHz and an amplitude of 0.5 mm to form sufficient sound pressure to manipulate cells without causing damage to the cells. By adjusting the frequency and amplitude of the acoustic wave generator, operations such as cell aggregation, dispersion, translation and rotation can be achieved. The integration of the acoustic wave generator with the microfluidic chip is achieved through precise alignment and connection to ensure that the acoustic wave energy is effectively transferred to the liquid medium. During the cell culture process, the output power of the acoustic wave generator can be adjusted in the range of 0.1 to 1 watt to meet the manipulation requirements of different cell types. By adjusting the frequency of the acoustic wave generator to 10 MHz and setting the amplitude to 0.5 mm, the aggregation operation of cell clusters can be achieved. This acoustic wave manipulation technique provides a non-invasive, high-throughput method for cell culture and analysis with broad application prospects.

[0039] The remote monitoring platform uses advanced network technology to achieve real-time monitoring of cell culture status. Users can access the platform anytime and anywhere through mobile devices such as smartphones or tablets. The platform supports encrypted data transmission through secure network protocols such as HTTPS to ensure data security and privacy. Users can view images and data of cell culture in real time through dedicated applications on mobile devices, including key parameters such as cell density, growth rate, and chemical composition of culture medium. The user interface design of the remote monitoring platform is intuitive and easy to use. Researchers can remotely control the cell culture environment through simple operations, such as adjusting temperature, pH value, and gas concentration. The platform also provides an alarm system. When cell culture conditions exceed the preset range, the system will automatically send notifications to users to ensure timely measures. For example, the system can set the temperature alarm threshold to 37℃±0.5℃ and the pH alarm threshold to 7.4±0.1 to ensure the stability of the cell culture environment. In addition, the remote monitoring platform can also store and analyze long-term accumulated cell culture data to help researchers optimize experimental plans and improve experimental efficiency. Through big data analysis and machine learning algorithms, the platform can predict cell growth trends and culture effects, providing strong data support for scientific research. This remote monitoring and management capability allows researchers to access and control the cell culture system through the Internet at any location, view the cell culture status in real time, adjust the culture parameters, and even replace the chip unit remotely when necessary, greatly improving the convenience and flexibility of the system. Through the efficient integration and intelligent connection of these modules, the system of the present invention ensures the automation, accuracy and environmental stability of the cell culture process, providing a powerful tool for biomedical research and drug development.

[0040] The 3D cell culture module is able to construct complex tissue models to simulate the environment of real biological tissues through its multi-layer structural design. The module uses an innovative multi-layer hydrogel technology, in which the thickness of each layer of hydrogel can be controlled between 100 microns and 500 microns to adapt to the attachment and growth needs of different cell types. By precisely controlling the hydrogel composition and porosity of each layer, a microenvironment with specific mechanical properties and biochemical signals can be created to promote cell proliferation and differentiation. For example, the porosity of the hydrogel can be adjusted between 85% and 95% to ensure the effective transmission of oxygen and nutrients to cells in 3D culture. In the process of constructing tissue models, the 3D cell culture module can achieve a gradient distribution of oxygen, nutrients, metabolites and soluble signals to form a diverse cell population. For example, by adjusting the oxygen permeability in the hydrogel, the oxygen gradient of different tissues in the body can be simulated, thereby affecting the behavior and function of cells. In addition, the module is also able to simulate cell-to-cell interactions, including three-dimensional cell-to-cell contacts and direct or indirect cell-to-cell communication, which is crucial for studying the behavior of cells in vivo. In order to achieve these complex 3D structures, the 3D cell culture module uses a combination of multiple cell types and biomaterials. For example, a natural ECM gel derived from the basement membrane of mouse sarcoma cells can be used, and its main components include laminin, type IV collagen, proteoglycan heparan sulfate and nestin, which can provide rich nutrition and are highly compatible with cell development. By planting cells in these prefabricated hydrogel layers, tissue models with specific structures and functions can be formed. Finally, through the automated operation module, researchers can easily replace the 3D cell culture module to adapt to different experimental needs. This modular design not only improves the flexibility of the experiment, but also simplifies the experimental operation, making the 3D cell culture technology more efficient and user-friendly. Through these innovative designs and functions, the system of the present invention provides a powerful tool for biomedical research and drug development, especially in the fields of tissue engineering and regenerative medicine, showing great application potential.

[0041] The self-circulating cooling system significantly improves the temperature control capability by integrating thermoelectric modules. Thermoelectric modules, also known as Peltier elements, use the Peltier effect to achieve heat movement. Depending on the direction of the current, one side absorbs heat while the other side releases heat, thereby achieving precise temperature control. This module can quickly adjust the temperature according to the specific needs of cell culture to ensure the stability of the cell culture environment. The design specifications of the thermoelectric module can be customized to meet different cell culture needs. For example, the size of the module can range from 20mm to 40mm to accommodate culture chambers of different sizes. The thermoelectric module can be built-in with a temperature sensor, such as a thermistor, to achieve high-precision temperature control with an accuracy of ±0.05℃. In addition, a heat sink or heat sink can be connected to the surface of the module to enhance the heat dissipation effect, which is essential for maintaining a constant temperature during cell culture. During the cell culture process, the thermoelectric module can work in conjunction with the heat sink and temperature sensor of the microfluidic chip to monitor and adjust the temperature of the chip in real time through the control of the automated operation module. When the chip temperature exceeds the preset range, the thermoelectric module will automatically start, and by adjusting the direction and size of the current, the chip temperature will be quickly adjusted back to a range suitable for cell growth, such as maintaining a physiological temperature of 37°C ± 0.1°C. In addition, the application of thermoelectric modules is not limited to temperature control, but can also be used for thermal management in the cell culture process, such as maintaining a constant temperature environment in a cell culture incubator, or providing necessary thermal cycles during cell culture. This modular design allows the thermoelectric modules to be quickly replaced and adjusted according to different experimental requirements, thereby improving the flexibility and adaptability of the cell culture system. Through the integration of thermoelectric modules, the automated microfluidic cell culture system of the present invention can achieve precise control of the cell culture environment, providing a stable and controllable temperature condition for cell culture, thereby improving the efficiency and quality of cell culture. The design of this self-circulating heat dissipation system not only enhances the temperature control capability, but also provides new possibilities for the development of cell culture technology.

[0042] The system of the present invention also includes environmentally friendly material design and energy-saving design process, and the sustainability of the system is significantly improved by integrating solar panels. These solar panels are designed to match the energy needs of the system, providing renewable energy and reducing dependence on traditional power grids. The peak power output of the solar panels is set to 300W, which is sufficient to meet the energy needs of the system under sufficient sunlight conditions. In the system design, the size and angle of the solar panels are optimized to maximize the absorption of solar energy while taking into account the absence of shadows or reflections on the surrounding environment. For example, the size of the solar panel can be 1.65 meters x 0.991 meters, and the angle is adjusted to the local latitude plus 5°-10° to ensure optimal energy collection efficiency. The solar panels are connected to other components of the system through an efficient energy management system that can intelligently distribute and regulate energy to ensure the continuity and stability of the cell culture process. In the absence of sunlight or at night, the system automatically switches to backup energy, such as battery storage or grid power supply, to ensure uninterrupted cell culture. This design not only improves energy efficiency, but also reduces the carbon footprint of the system, which is in line with the concept of green building and sustainable development. The integration of solar panels also helps to reduce the carbon footprint of the system, which is in line with the concept of green building and sustainable development. Through this design, the system not only improves energy efficiency, but also demonstrates the possibility of implementing environmental protection and energy-saving measures in biomedical research facilities. This innovative environmentally friendly material and energy-saving design make the automated microfluidic cell culture system a model of environmental protection and energy saving while providing high-precision cell culture.

[0043] The present invention provides a method for cell culture using an automated microfluidic cell culture system integrated with artificial intelligence, comprising the following steps: S1. Initialize the microfluidic chip: First, install the microfluidic chip into the system and sterilize it with ultraviolet radiation for 30 minutes to ensure the sterility of the chip surface. Then, inject the basal culture medium preheated to 37°C into the microfluidic channel of the chip through the precision pump of the automated operation module, and control the flow rate at 0.1 mL / min to ensure that the channel is fully infiltrated and free of bubbles.

[0044] S2. Precise distribution of cells or microspheres: Using digital microfluidics technology, cells or microspheres are precisely distributed to specific areas of the microfluidic chip at a flow rate of 0.05 mL / min through electrowetting or dielectrophoresis drive mechanisms. Approximately 10,000 cells are inoculated in each chamber to ensure uniform cell distribution.

[0045] S3. Automatic culture medium replacement and nutrient addition: Through the automated operation module, the cell culture medium is automatically replaced, nutrients are added, and waste is removed. The culture medium replacement frequency is set to once every 24 hours, and nutrient addition is adjusted according to cell growth needs, usually once every 48 hours.

[0046] S4. Monitor and adjust the culture environment: Use an artificial intelligence control unit to monitor the cell culture environment, including maintaining the temperature at 37℃±0.1℃, controlling the pH value at 7.4±0.05, and setting the oxygen concentration to 5% CO2. These parameters are monitored in real time by sensors and automatically adjusted to the set values.

[0047] S5. Remote observation and operation: Through the remote monitoring platform, researchers can observe the cell status in real time, including cell morphology, growth rate and cell density, and perform necessary operations. The platform provides images and data updated every 10 minutes to ensure that researchers can respond in time.

[0048] S6. Construction and cultivation of organoid models: In the 3D cell culture module, scaffolds and biomaterials are used to construct three-dimensional cell structures to simulate complex biological tissue structures. The temperature of the module is maintained at 37°C, the pH value is controlled at 7.2, and the oxygen concentration is 20% O2 to simulate the in vivo environment.

[0049] S7. Maintain constant temperature: The self-circulating cooling system uses a microfluidic heat sink and a temperature sensor to maintain a constant temperature of the chip. The thermal conductivity of the heat sink is set to 200 W / mK to ensure rapid heat dissipation.

[0050] S8. Sample collection and immediate analysis: Samples are collected and analyzed regularly during the cell culture process through an automated operation module to evaluate the cell culture effect. The sample collection cycle is once every 72 hours, and the analysis includes cell viability, metabolic activity, and protein expression levels.

[0051] S9. Data analysis and culture program optimization: The AI ​​control unit is used to analyze the collected data and optimize the subsequent cell culture program. The analysis results are used to adjust the culture medium formula, nutrient concentration and culture conditions.

[0052] S10. Collecting cultures: After cell culture is completed, collect the cultures, including cell layers, cell spheres, or organoid models, through the automated operation module. During the collection process, ensure aseptic operation to avoid cell contamination.

[0053] S11. Subsequent analysis: The collected cultures are subjected to subsequent biological, pharmacological or toxicological analysis to evaluate their function and safety. The analysis includes gene expression analysis of cells, drug response testing and toxicity assessment to ensure the quality and suitability of the cultures.

[0054] Through the above steps, the automated microfluidic cell culture system of the present invention can provide a standardized, automated cell culture solution, which is applicable to multiple fields such as biomedical research, drug screening, tissue engineering and disease model research.

[0055] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and descriptions are only preferred examples of the present invention and are not intended to limit the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. An automated microfluidic cell culture system integrating artificial intelligence, characterized in that: include: Microfluidic chip, artificial intelligence control unit, automated operation module, digital microfluidic module, remote monitoring platform, 3D cell culture module, self-circulating cooling system; among them, The microfluidic chip is the core of the system, integrating microfluidic channels, reaction chambers, gas exchange areas, and waste collection systems for cell culture to provide a growth environment for cells; The artificial intelligence control unit is connected to the microfluidic chip, analyzes the cell culture data from the microfluidic chip through an integrated machine learning algorithm, and automatically adjusts the culture conditions in real time through the automated operation module; An automated operation module that interfaces with the microfluidic chip through precision pumps, valves, temperature controllers, and gas supply systems to perform precise liquid delivery and environmental control operations; Digital microfluidic modules, combined with the microfluidic channels of microfluidic chips, use electrowetting or dielectrophoresis drive mechanisms to achieve non-contact manipulation of cells; A remote monitoring platform, connected to the AI ​​control unit via a wireless or wired network, provides a user interface and data management system, allowing researchers to monitor and record cell culture parameters and historical data in real time; 3D cell culture module, combined with microfluidic chip, uses scaffolds and biomaterials to form three-dimensional cell structures within the microfluidic chip; The self-circulating cooling system is connected to the microfluidic chip through a microfluidic heat sink and a temperature sensor to maintain a constant temperature of the microfluidic chip.

2. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: It also includes a power management module that collects solar energy through solar panels to power the entire system.

3. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: The microfluidic chip has a modular design, including interchangeable chip units, to accommodate different cell culture requirements.

4. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: The artificial intelligence control unit includes image recognition software for analyzing cell morphology and growth status.

5. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: The automated operation module includes an automatic sampler and an analyzer, which are used for sample collection and instant analysis during the cell culture process.

6. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: The digital microfluidic module also includes an acoustic wave generator for assisting the manipulation of cells or microspheres.

7. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: The 3D cell culture module includes a multi-layer structure and is used to construct a complex tissue model.

8. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: The self-circulating heat dissipation system also includes a thermoelectric module for enhancing the temperature control capability.

9. The automated microfluidic cell culture system integrated with artificial intelligence according to claim 1, characterized in that: The system also includes environmentally friendly material design and energy-saving design process.

10. An automated microfluidic cell culture method with integrated artificial intelligence based on an automated microfluidic cell culture system with integrated artificial intelligence according to any one of claims 1 to 9, characterized in that: The steps include: S1, initializing the microfluidic chip and configuring the microfluidic channels and reaction chambers required for cell culture; S2, using a digital microfluidics module to accurately distribute cells or microspheres to predetermined areas of a microfluidics chip; S3, automatically replacing cell culture medium, adding nutrients and removing waste through the automated operation module; S4, using an artificial intelligence control unit to monitor the cell culture environment and adjust the culture conditions in real time; S5. Remotely observe cell status, including cell morphology, growth rate and cell density, through a remote monitoring platform; S6. Build and culture organoid models in 3D cell culture modules to simulate complex biological tissue structures; S7. Use a self-circulating cooling system to maintain a constant temperature during cell culture to ensure that cell growth is not affected by temperature fluctuations; S8. Regularly collect samples and conduct instant analysis during the cell culture process through the automated operation module to evaluate the cell culture effect; S9, using the artificial intelligence control unit to analyze the collected data and optimize the subsequent cell culture program; S10. After the cell culture is completed, the culture is collected through an automated operation module, including a cell layer, a cell sphere or an organoid model; S11. Perform subsequent biological, pharmacological, or toxicological analyses on the collected cultures to evaluate their functionality and safety.

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