A cell culture metabolite real-time detection device

Abstract

The application discloses a real-time detection device for cell culture metabolites. Compared with the prior art, the application guarantees the overall structure of the device to be heavy at the bottom and light at the top by adding a lithium battery and a counterweight in the device, so that the orientation of the device when placed in water is guaranteed, so that the detection end of the electrochemical biosensor can always be below the liquid level. And the application configures an antenna module connected with the electrochemical collection module in the device, so that it can display and transmit experimental data in real time, and guarantee the real-time feedback of experimental data.

Description

Technical Field

[0001] This application relates to the field of cell culture, and in particular to a real-time detection device for cell culture metabolites. Background Technology

[0002] During cell culture, the dynamic changes in metabolites (such as glucose, lactic acid, glutamine, and oxygen) reflect the physiological state, metabolic level, and response to external stimuli of cells. These are key parameters for experiments in cell biology, drug screening, and disease model research. Therefore, real-time and continuous monitoring of metabolite concentrations in cell culture medium is of great significance for obtaining firsthand information on changes in cell state, finely controlling culture conditions, and improving experimental reproducibility and accuracy.

[0003] Currently, mainstream methods for detecting cellular metabolites include mass spectrometry (MS), chromatography (such as HPLC and GC), and conventional biochemical detection methods. While these methods offer good sensitivity and accuracy, they typically rely on offline sample acquisition, processing, and analysis, resulting in significant limitations such as cumbersome operation, long processing times, and difficulty in real-time monitoring. Furthermore, these methods require sophisticated equipment when processing trace samples, are susceptible to sample contamination, and cannot continuously provide metabolic information, making it difficult to reflect dynamic changes in cell state.

[0004] In recent years, some studies have attempted to extract culture medium using microfluidic chips or automated systems, and then connect sensors or analytical devices for detection. However, these systems are often complex in structure, rely on external pumps or tubing, pose potential contamination risks, and the extraction process itself may disturb the cell growth environment. Furthermore, this intermittent extraction still cannot meet the need for high temporal resolution monitoring of rapid metabolic processes.

[0005] Therefore, there is an urgent need in this field for an integrated spherical real-time monitoring device that can directly detect the concentration of target metabolites in liquids. Summary of the Invention

[0006] The purpose of this application is to provide an integrated spherical real-time monitoring device: the lower part of the device is an electrochemical biosensor module that can be in contact with the culture medium for a long time, used to directly detect the concentration of target metabolites in the liquid, and it can automatically measure the real-time metabolite curve during cell culture, thereby ensuring the real-time feedback of experimental data.

[0007] This application discloses a real-time detection device for cell culture metabolites, the device comprising: a bottom, a shell, an electrochemical acquisition module, an antenna module, and a battery module, wherein:

[0008] The housing is detachably connected to the bottom, and the housing has openings on the top and one side;

[0009] The electrochemical acquisition module is disposed at the bottom of the housing and includes an electrochemical biosensor, an electrode connector, and an electrochemical data acquisition unit. One end of the electrochemical biosensor is connected to the battery module through the electrode connector and to the electrochemical data acquisition unit through the acquisition circuit. The other end extends out of the lower part of the housing and passes through a pre-reserved opening in the bottom to ensure that it is submerged in liquid. The electrochemical biosensor is configured to send the acquired data into the electrochemical data acquisition unit.

[0010] The antenna module is disposed within the housing and includes an antenna connected to the acquisition circuit and extending from an opening at the top of the housing; the antenna module is configured to receive data from the electrochemical data acquisition unit and transmit it to the outside.

[0011] The battery module includes a lithium battery, a wireless charging coil, and a wireless charger; the lithium battery is connected to the electrochemical biosensor via the electrode connector, and the wireless charging coil is disposed on the lithium battery near the side opening of the housing; the wireless charger is configured to connect to the wireless charging coil via the side opening of the housing.

[0012] In a preferred embodiment, the connection between the housing and the bottom is configured as a closed connection, the closed connection comprising sealing with a sealing ring or sealant.

[0013] In a preferred embodiment, the bottom interior is further provided with a buoyancy-filling material; the buoyancy-filling material is configured to ensure that the liquid level does not exceed the junction between the outer shell and the bottom.

[0014] In a preferred embodiment, the electrochemical biosensor is a three-electrode system designed based on micro / nano fabrication technology.

[0015] In a preferred embodiment, the number of electrochemical biosensors is not less than three.

[0016] In a preferred embodiment, a counterweight is disposed in the bottom to ensure the orientation of the detection device.

[0017] In a preferred embodiment, the electrochemical biosensor is detachably configured within the detection device.

[0018] In a preferred embodiment, the outer casing and the bottom are snap-fit ​​connected.

[0019] In a preferred embodiment, the overall diameter of the detection device is 3-7 cm.

[0020] In a preferred embodiment, each of the plurality of said electrochemical biosensors is configured to monitor different biological data.

[0021] In a preferred embodiment, an isolation plate is provided at the connection between the bottom and the outer casing, and the isolation plate is provided with an opening corresponding to the bottom opening of the bottom.

[0022] In a preferred embodiment, the size of the buoyancy material is larger than the opening.

[0023] In a preferred embodiment, the counterweight is fixed to the bottom and has an opening.

[0024] The advantages of this invention include:

[0025] (1) This invention adopts a modular design, which facilitates maintenance and reuse: the device adopts an upper and lower detachable structure, in which the electrochemical biosensor module can be replaced independently. The performance of the electrochemical biosensor will degrade after prolonged use, making it easy to replace aging sensor components. The upper control module can be reused for a long time, saving experimental costs and extending the service life of the equipment.

[0026] (2) This invention has no external wiring or pipelines, and is easy to operate: The entire device is designed to be completely wireless and without external pipeline connections, avoiding problems such as pollution and blockage caused by pipelines or pump systems in traditional sampling systems, significantly reducing the risk of bacterial growth and cross-contamination, and simplifying the experimental process.

[0027] (3) This invention achieves strong compatibility and adaptability to various experimental environments by setting the detection content of the electrochemical sensor. Furthermore, the device is small in size and compactly packaged, and can be directly placed into various cell culture containers, including multi-well plates, culture dishes, shake flasks, bioreactors, etc., without requiring additional modifications to existing experimental systems, making it easy to promote and apply.

[0028] (4) Low power consumption design to support long-term monitoring: The internal electronic system adopts low power chips and optimized power management scheme, and with the help of rechargeable lithium battery, it can run continuously for a long time to meet the long-term metabolic monitoring needs in the cell culture process.

[0029] (5) High scalability: The sensor module can be replaced with different types of electrochemical sensors according to actual needs, so as to realize customized detection of a variety of metabolites (such as glucose, lactic acid, glutamine, oxygen, pH, etc.) and is suitable for different research scenarios.

[0030] (6) Minimal interference with the culture environment: The device is used in an immersion manner, with no bubbles, vibrations, or liquid disturbances during operation, which has minimal impact on the cell growth environment. It is especially suitable for organoids, 3D culture systems, or stem cell culture systems that are sensitive to environmental changes.

[0031] The specification of this application contains numerous technical features distributed across various technical solutions. Listing all possible combinations of these technical features (i.e., technical solutions) would make the specification excessively lengthy. To avoid this problem, the various technical features disclosed in the above-described invention, the various technical features disclosed in the following embodiments and examples, and the various technical features disclosed in the accompanying drawings can be freely combined to form various new technical solutions (all of which should be considered as described in this specification), unless such a combination of technical features is technically infeasible. For example, one example discloses feature A+B+C, and another example discloses feature A+B+D+E. Features C and D are equivalent technical means that serve the same function, and technically only one needs to be used; they cannot be used simultaneously. Feature E can technically be combined with feature C. Therefore, the solution A+B+C+D should not be considered as described because it is technically infeasible, while the solution A+B+C+E should be considered as described. Attached Figure Description

[0032] Figure 1 This is a schematic diagram of the overall structure of this application;

[0033] Figure 2 This is a circuit connection diagram of this application;

[0034] Figure 3 This is a schematic diagram of the mechanical structure of this application;

[0035] Figure 4 This is a schematic diagram of the mechanical structure of this application in the charging state; Figure 5 This is a schematic diagram of an embodiment of the present application, in which a counterweight block is configured at the bottom and buoyancy material is filled.

[0036] Explanation of reference numerals in the attached figures:

[0037] 1-Outer shell; 2-Bottom; 3-Buoyancy filling material; 4-Electrochemical biosensor; 5-Electrode connector; 6-Lithium battery; 7-Electrochemical data acquisition unit; 8-Wireless charging coil; 9-Wireless charger; 10-RF antenna; 11-Counterweight; 12-Isolation plate; 13-Snap fastener. Detailed Implementation

[0038] Through in-depth research and extensive screening, the inventors of this application have developed a real-time detection device for cell culture metabolites. Compared with existing technologies, this application, by incorporating a lithium battery and a counterweight, ensures a bottom-heavy, top-light structure, thereby guaranteeing the device's orientation when placed in water and ensuring that the detection end of the electrochemical biosensor remains below the liquid surface. Furthermore, by configuring an antenna module connected to the electrochemical acquisition module within the device, this application enables real-time display and transmission of experimental data, ensuring timely feedback of experimental data.

[0039] To make the objectives, technical solutions, and advantages of this application clearer, the embodiments of this application will be described in further detail below with reference to the accompanying drawings.

[0040] Example 1

[0041] The real-time detection device for cell culture metabolites of the present invention, such as Figure 1 , Figure 3 and Figure 4 As shown, the real-time monitoring device includes: a bottom 2, a housing 1, an electrochemical acquisition module, an antenna module, and a battery module, wherein:

[0042] The outer casing 1 is detachably connected to the bottom 2, and the outer casing 1 has openings on its top and one side; in this embodiment, as... Figure 3 As shown, the outer shell 1 and the bottom 2 are combined to form an elliptical structure, with the bottom 2 occupying one-quarter of the bottom of the ellipse and the remainder being the outer shell 1.

[0043] Optionally, in one embodiment, the bottom 2 is further provided with a buoyancy material 3; the buoyancy material 3 is configured to ensure that the liquid level does not exceed the junction of the outer shell and the bottom. Optionally, in another embodiment, the bottom 2 is provided with a counterweight 11 to ensure the orientation of the device when it is in water.

[0044] Optionally, in one embodiment, the outer shell 1 and the bottom 2 are connected by a snap-fit, and the connection between the outer shell 1 and the bottom 2 is sealed by using a sealing ring or sealant.

[0045] Optionally, in one embodiment, a partition plate 12 is disposed between the outer casing 1 and the bottom 2.

[0046] The electrochemical acquisition module is disposed at the bottom of the housing and includes an electrochemical biosensor 4, an electrode connector 5, and an electrochemical data acquisition unit 7. One end of the electrochemical biosensor 4 is connected to the battery module and the electrochemical data acquisition unit 7 via the electrode connector 5, while the other end extends out from the lower part of the housing 1 and passes through a pre-reserved opening in the bottom 2, ensuring it is submerged in liquid. The electrochemical biosensor 4 is configured to transmit the acquired data to the electrochemical data acquisition unit 7. In this embodiment, the electrochemical biosensor is a three-electrode system designed based on micro / nano fabrication technology. Optionally, in one embodiment, the number of electrochemical biosensors 4 is 3-7, and they are detachably connected to the electrode connector 5, ensuring immediate replacement if any electrochemical biosensor 4 is damaged. Alternatively, multiple types of electrochemical biosensors 4 can be configured to perform real-time measurement of different data.

[0047] The antenna module is disposed within the housing and includes a radio frequency antenna 10 extending from an opening at the top of the housing; the antenna module is configured to receive data from the electrochemical data acquisition unit 7 and transmit it to the outside.

[0048] The battery module includes a lithium battery 6, a wireless charging coil 8, and a wireless charger 9; the lithium battery 6 is connected to the electrochemical biosensor 4 through the electrode connector 5, and the wireless charging coil 8 is disposed on the lithium battery near the side opening of the housing; the wireless charger 9 is configured to connect to the wireless charging coil 8 through the side opening of the housing 1.

[0049] Example 2

[0050] This embodiment discloses a circuit structure diagram of the device of the present invention, specifically, as follows: Figure 2 As shown, the electrochemical sensor 4 performs data acquisition, the battery module powers the electrochemical data acquisition unit 7, and the information received by the electrochemical data acquisition unit 7 is sent to the antenna module. (Further details omitted) Figure 1 The data is transmitted to an external system via the antenna module, thereby enabling real-time data monitoring during the biological culture process.

[0051] Example 3

[0052] Another embodiment of the invention, for example Figure 5 As shown, the real-time monitoring device includes: a bottom 2, a housing 1, an electrochemical acquisition module, an antenna module, and a battery module, wherein:

[0053] The outer casing 1 and the bottom 2 are detachably connected by a snap fastener 13, and the outer casing 1 has openings on its top and one side; in this embodiment, as Figure 4As shown, the bottom 2 and the bottom 2 are combined to form an elliptical structure, with the bottom 2 occupying half the volume of the ellipse.

[0054] The electrochemical data acquisition module is located in the middle of the entire device, integrated within and extending out of the outer casing 1. It includes an electrochemical biosensor 4, an electrode connector 5, and an electrochemical data acquisition unit 7. One end of the electrochemical biosensor 4 is connected to the battery module and the electrochemical data acquisition unit 7 via the electrode connector 5, while the other end extends out from the lower part of the outer casing 1 and passes through a pre-reserved opening in the bottom 2, ensuring it is submerged in liquid. The electrochemical biosensor 4 is configured to transmit the acquired data to the electrochemical data acquisition unit 7. In this embodiment, the electrochemical biosensor is a three-electrode system designed based on micro-nano fabrication technology, and the number of electrochemical sensors is five. It is detachably connected to the electrode connector 5, ensuring immediate replacement if any electrochemical biosensor 4 is damaged. Alternatively, multiple types of electrochemical biosensors 4 can be configured to perform real-time measurements of different data.

[0055] The antenna module is disposed within the housing and includes a radio frequency antenna 10 extending from an opening at the top of the housing; the antenna module is configured to receive data from the electrochemical data acquisition unit 7 and transmit it to the outside.

[0056] The battery module includes a lithium battery 6, a wireless charging coil 8, and a wireless charger 9; the lithium battery 6 is connected to the electrochemical biosensor 4 through the electrode connector 5, and the wireless charging coil 8 is disposed on the lithium battery near the side opening of the housing; the wireless charger 9 is configured to connect to the wireless charging coil 8 through the side opening of the housing 1.

[0057] In this embodiment, a counterweight 11 and buoyancy material 3 are disposed at the bottom. To prevent the counterweight 11 and buoyancy material 3 from entering the outer shell during placement or storage, an insulating plate 12 is disposed at the joint between the outer shell 1 and the bottom 2. The insulating plate 12 has an opening corresponding to the bottom 2 for the electrochemical data sensor to pass through. In this embodiment, the volume of the buoyancy material 3 is much larger than the opening to prevent it from passing through. The counterweight 11 is also disposed on the insulating plate 12 with an opening. Optionally, in one embodiment, there are multiple counterweights 11, each with a small volume but larger than the opening, so that they do not need to be fixed to the insulating plate.

[0058] It should be noted that in this patent application, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one" does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element. In this patent application, if it refers to performing an action according to an element, it means performing the action at least according to that element, including two cases: performing the action only according to that element, and performing the action according to that element and other elements. Expressions such as "multiple," "repeatedly," and "various" include two, two times, two kinds, and more than two, more than two times, and more than two kinds.

[0059] This specification includes combinations of various embodiments described herein. Individual references to “one embodiment” or a particular embodiment, etc., do not necessarily refer to the same embodiment; however, these embodiments are not mutually exclusive unless indicated to be mutually exclusive or are readily apparent to those skilled in the art. It should be noted that the word “or” is used in a non-exclusive sense throughout this specification unless the context explicitly indicates or requires it.

[0060] All references to this application are considered to be incorporated herein by reference in their entirety so as to serve as a basis for modification if necessary. Furthermore, it should be understood that after reading the foregoing disclosure of this application, those skilled in the art can make various alterations or modifications to this application, and these equivalent forms also fall within the scope of protection claimed in this application.

Claims

1. A real-time detection device for cell culture metabolites, characterized in that, The device includes: a bottom, a housing, an electrochemical acquisition module, an antenna module, and a battery module, wherein: The housing is detachably connected to the bottom, and the housing has openings on the top and one side; The electrochemical acquisition module is disposed at the bottom of the housing and includes an electrochemical biosensor, an electrode connector, and an electrochemical data acquisition unit. One end of the electrochemical biosensor is connected to the battery module through the electrode connector and to the electrochemical data acquisition unit through the acquisition circuit. The other end extends out of the lower part of the housing and passes through a pre-reserved opening in the bottom to ensure that it is submerged in liquid. The electrochemical biosensor is configured to send the acquired data into the electrochemical data acquisition unit. The antenna module is disposed within the housing and includes an antenna connected to the acquisition circuit and extending from an opening at the top of the housing; the antenna module is configured to receive data from the electrochemical data acquisition unit and transmit it to the outside. The battery module includes a lithium battery, a wireless charging coil, and a wireless charger; the lithium battery is connected to the electrochemical biosensor via the electrode connector, and the wireless charging coil is disposed on the lithium battery near the side opening of the housing; the wireless charger is configured to connect to the wireless charging coil via the side opening of the housing.

2. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, The connection between the outer casing and the bottom is configured as a closed connection, which includes sealing using a sealing ring or sealant.

3. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, The bottom is also filled with buoyancy material; the buoyancy material is configured to ensure that it does not exceed the joint between the outer shell and the bottom.

4. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, The electrochemical biosensor is a three-electrode system designed based on micro-nano fabrication technology.

5. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, The number of electrochemical biosensors shall not be less than three.

6. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, A counterweight is provided at the bottom to ensure the orientation of the detection device.

7. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, The electrochemical biosensor is detachably configured within the detection device.

8. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, The outer shell and the bottom are connected by a snap-fit.

9. The real-time detection device for cell culture metabolites according to claim 1, characterized in that, The overall diameter of the detection device is 3-7 cm.

10. The cell culture metabolite real-time detection device according to claim 1, wherein each of the plurality of electrochemical biosensors is configured to monitor different biological data.

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