Living cell tissue perfusion collection system

Abstract

The utility model application relates to the field of biological experiment equipment discloses a kind of living cell tissue perfusion collection systems.The system includes: liquid flow switching dosing module, constant temperature incubation module, sample automatic collection module and sample low-temperature maintenance module.Liquid flow switching dosing module switches different liquid according to experimental requirement anytime, average shunt pumps into constant temperature incubation module;Constant temperature incubation module provides closed, constant temperature incubation liquid flow environment for living cell tissue perfusion experiment;Sample automatic collection module receives the perfusion liquid sample flowing through constant temperature incubation module, and according to the specified time sequence, sample is collected into collection well plate;Sample low-temperature maintenance module keeps sample collection plate in relatively low temperature state by refrigeration temperature control mode, to delay sample volatilization and the denaturation of the substance to be tested in it, guarantee sample stability.Through the system, perfusion liquid sample can be accurately collected according to time sequence, improve experimental result accuracy, to obtain repeatable high-quality experimental result.

Description

Technical Field

[0001] This application relates to the field of biological experimental equipment, specifically to a live cell tissue perfusion collection system. Background Technology

[0002] Biomedical research is one of the most watched scientific research fields globally today, and its development is inseparable from the continuous advancement of experimental techniques. Modern biomedical experimental techniques, such as mass spectrometry, laser imaging, in vivo mouse imaging, live cell and animal metabolic monitoring, and gene analysis, have greatly promoted the in-depth development of physiological mechanism research. Currently, international biomedical research on metabolic mechanisms mainly focuses on the following two directions:

[0003] 1. Tracking and analysis of metabolism-related molecules: Using mass spectrometry and nuclear magnetic resonance (NMR) technology, the metabolic molecules such as sugars, fats, proteins, vitamins, macro- and micro-elements can be quantitatively analyzed and their metabolic pathways in organisms can be tracked, such as the absorption, metabolism and excretion of glucose and fatty acids.

[0004] 2. Research on regulatory factors in metabolic processes: Focus on the role of small molecules or biological macromolecules (such as insulin) in metabolic processes and metabolic diseases, especially the metabolites secreted by secretory cells (such as adipocytes, chromaffin cells, hepatocytes, etc.) under specific stimuli, which has important research value.

[0005] The realization of the above research directions relies on the measurement of metabolic parameters of living cells or tissues, especially the collection of cellular response products and secretions. Real-time, quantitative sample collection would significantly improve experimental quality and research efficiency. Traditional methods typically employ static culture dishes or ex vivo tissue baths, where the culture medium does not flow, affecting the continuity and reproducibility of experiments. Subsequently, unidirectional flow microfluidic experimental devices and ex vivo tissue baths were developed; however, the former is limited in experimental scale and difficult to reuse, while the latter has a large volume, making it unsuitable for small-sample tissue experiments. Existing technologies also suffer from problems such as obstructed fluid flow, drug residue, droplet splashing, and sample evaporation, affecting experimental stability and data reproducibility. Summary of the Invention

[0006] The purpose of this application is to provide a biological experimental device that can accurately and in real time collect perfusion fluid samples, improve the accuracy of experimental results, and thus obtain reproducible high-quality experimental results.

[0007] This application discloses a live cell tissue perfusion collection system, including: a fluid flow switching drug delivery module, a constant temperature incubation module, an automatic sample collection module, and a sample low temperature maintenance module.

[0008] The fluid flow switching drug delivery module includes a fluid flow dynamic structure and a fluid flow switching and dispensing structure; it is configured to switch, dispense, and pump different liquids into the constant temperature incubation module at any time according to experimental requirements.

[0009] The isothermal incubation module is configured to provide a closed, isothermal incubation fluid environment for live cell tissue perfusion experiments.

[0010] The automatic sample collection module is configured to receive liquid samples flowing through the isothermal incubation module and collect the samples into the collection well plate according to a specified time sequence.

[0011] The sample low-temperature maintenance module keeps the sample collection plate at a relatively low temperature through water baths, metal baths, etc., in order to delay sample volatilization and denaturation of the analytes, and ensure sample stability.

[0012] During operation, the live cell tissue perfusion collection system utilizes a fluid switching drug delivery module to pump different perfusion fluids into the incubation chamber of the constant temperature incubation module after switching and separating them at different times. After perfusing the live cell tissue in the incubation chamber, the fluid flows into the collection wells of the collection plate of the automatic sample collection module. The constant temperature incubation module maintains a suitable survival temperature for the cells to preserve their viability, while the low temperature sample maintenance module lowers and maintains the temperature at a sufficiently low level to ensure the stability of the activity of the liquid sample collected by the collection plate.

[0013] In a preferred embodiment, the live cell tissue perfusion collection system further includes a fluid flow switching drug delivery module, which comprises a fluid flow dynamic structure and a fluid flow switching and dispensing structure. The fluid flow dynamic structure is configured to provide power for the fluid flow during the live cell tissue perfusion experiment, ensuring the fluid flows within a closed pipeline. The fluid flow dynamic structure is designed to quantitatively control the speed, time, and dosage. Depending on the experimental requirements for quantitative control of fluid flow and time, multi-channel or single-channel peristaltic pumps, diaphragm pumps, micro-injection pumps, independent multi-channel micro-injection pump systems, etc., can be selected to deliver the fluid flow through the tubing.

[0014] The fluid switching and dispensing structure of the fluid switching drug delivery module is composed of at least, but not limited to, a multi-channel switching valve, a solenoid valve, a multi-channel dispenser, and combinations thereof. Depending on different experimental setup requirements, one or more components may be used to input various perfusion liquids through multiple inlets and distribute them evenly to multiple incubation chambers of the constant temperature incubation module.

[0015] When the fluid dynamic structure is selected as a micro-injection pump combination or a multi-channel micro-injection pump, the micro-injection pump combination or the multi-channel micro-injection pump can undertake the functions of fluid flow switching and dispensing, which partially simplifies the structure and composition of the fluid flow switching and dispensing of the fluid flow switching drug delivery module.

[0016] In a preferred embodiment, the live cell tissue perfusion collection system further includes a constant temperature incubation module, which consists of multiple incubation chambers and a heating temperature control structure;

[0017] The incubation chamber is a closed cavity structure. Each incubation chamber contains the same number of live cells or ex vivo tissues and solid support materials. The upper end of the incubation chamber is provided with a perfusion fluid inlet. The perfusion fluid enters the multiple incubation chambers through the perfusion fluid inlet to perfuse the live cells or ex vivo tissues to provide nutrient supply or drug treatment. The lower end of the incubation chamber is provided with a perfusion fluid outlet. The outlet has a screen structure to ensure that the live cells or ex vivo tissues and solid support materials remain in the incubation chamber during the perfusion process. After the perfusion is completed, the perfusion fluid sample flows out from the perfusion fluid outlet to the automatic sample collection module. The perfusion fluid sample includes the metabolic products and secretions of the live cells or ex vivo tissues.

[0018] The heating and temperature control structure ensures a constant temperature within multiple incubation chambers, maintaining it within the range of room temperature to 45°C. Depending on the specific experimental setup requirements, various structures such as air baths, water baths, metal baths, and heating rods with temperature sensors can be used to maintain a constant temperature, generally controlled within the range of room temperature to 45°C.

[0019] In a preferred embodiment, the live cell tissue perfusion collection system further includes an automatic sample collection module, which consists of a liquid drainage structure and a sample collection time sequence control structure.

[0020] The liquid drainage structure is designed to ensure that droplets fall into the collection holes of the sample collection plate below, reducing sample liquid spillage outside the collection holes and minimizing the volume error of sample liquid collected in each collection hole. This structure can be, at least but not limited to, using a controllable, elastic fiber filament (drainage rod) that makes oblique contact and separation from the sample outlet and the sidewall of the collection hole; alternatively, a controllable support rod with a spring connecting a metal sheet (drainage plate) can make oblique contact and separation from the sample outlet and the sidewall of the collection hole; or the sample outlet can be obliquely positioned with a section of elastic silicone tube (drainage tube) at the front end. The relative position of the silicone tube end at the front end of the sample outlet and the sidewall of the collection hole on the sample collection plate can be controlled by movement, causing the silicone tube end at the front end of the sample outlet to make oblique contact and separation from the sidewall of the collection hole, etc., to ensure that droplets fall into the collection holes below, preventing sample liquid spillage outside the collection holes and minimizing the volume error of sample liquid collected in each collection hole.

[0021] The sample collection time sequence control structure is configured to receive liquid samples flowing through the isothermal incubation module and collect the samples into the collection well plate according to a specified time sequence. The sample collection time sequence control structure consists of a sample collection well plate, a fixed support plate, and a mechanical drive structure. The sample collection well plate is fixed to the fixed support plate, which is driven by the mechanical drive structure, which consists of a motor and a linear guide rail. During sampling, the fixed support plate, along with the sample collection well plate, performs horizontal linear displacement and pauses at certain time intervals, ensuring that the collection holes of the sample collection well plate are aligned with the perfusion fluid outlet of the incubation chamber above, guaranteeing that droplets fall into the collection holes sequentially according to time. The speed, movement time, and dwell time of the fixed support plate will be determined based on the experimental requirements for the time sequence, the volume of the collection holes on the well plate, and the required sample volume for subsequent experiments.

[0022] In a preferred embodiment, the live cell tissue perfusion collection system further includes a sample cryopreservation module. This module is configured to maintain the sample collection plate at a relatively low temperature using methods such as a water bath or metal bath to prevent liquid evaporation and condensation, while simultaneously delaying sample evaporation and denaturation of the analytes, thus ensuring sample stability. The refrigeration principle can be achieved through methods such as ice-water circulation, semiconductor refrigeration, or semiconductor refrigeration combined with refrigerant circulation.

[0023] In this embodiment, the fluid flow switching drug delivery module improves the fluid flow conversion and dispensing control structure and selects multiple fluid flow power sources according to specific experimental requirements, thus solving the contradiction between real-time drug delivery switching and pipeline liquid residue in the prior art and effectively avoiding cross-contamination and pipeline blockage between multiple perfusion fluids.

[0024] Furthermore, the constant temperature incubation module uses heating and insulation methods such as water baths and metal baths to make the ambient temperature for incubating perfused cells more constant, effectively reducing systematic errors between incubation chambers, and adopting a more sealed incubation chamber structure to avoid leakage of perfusion fluid.

[0025] Furthermore, the automatic sample collection module employs a liquid drainage structure to reduce the problem of sample liquid spilling outside the collection holes and to reduce the volume error of sample liquid in each collection hole. At the same time, the automatic sample collection module can be driven by its mechanical drive structure to make horizontal linear displacement at certain time intervals during the sampling process, ensuring that droplets fall into the collection holes accurately in time sequence, and can realize continuous sampling of multiple collection well plates, thus achieving the purpose of sampling according to time sequence.

[0026] Furthermore, the sample cryogenic maintenance module achieves low-temperature stability of the sample, prevents liquid sample evaporation and analyte denaturation, reduces the impact on experimental results, and improves the reliability of experimental data.

[0027] 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 are considered to have been 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

[0028] Figure 1 This is an overall schematic diagram of the device according to one embodiment of this application;

[0029] Figure 2 This is a schematic diagram of multi-channel drug delivery control switching according to one embodiment of this application;

[0030] Figure 3 This is a schematic diagram of a multi-channel dispensing drug delivery connection according to one embodiment of this application;

[0031] Figure 4 This is a schematic diagram of sample liquid collection and drainage according to one embodiment of this application;

[0032] Figure 5 This is a schematic diagram of a micro-injection pump or a multi-channel micro-injection pump as a fluid flow driving power source according to one embodiment of this application.

[0033] Explanation of reference numerals in the attached figures

[0034] 1. Liquid flow pipeline

[0035] 2. Source of perfusion fluid

[0036] 3. Liquid flow switching and liquid separation structure

[0037] 4. Hydrodynamic structure

[0038] 5. Heating temperature control structure (including air bath structure)

[0039] 6. Incubation chamber and support frame

[0040] 7. Liquid drainage structure

[0041] 8. Sample collection well plate

[0042] 9. Sample Low Temperature Maintenance Module

[0043] 10. Fixed support plate and mechanical drive structure

[0044] 11. Multi-channel switching valve or solenoid valve

[0045] 12. Multichannel dispenser

[0046] 13. Sample output port

[0047] 14. Droplets

[0048] 15. Drainage stick

[0049] 16. Collection Hole

[0050] 17. Drainage plate

[0051] 18. Spring

[0052] 19. Support rod

[0053] 20. Drainage tube

[0054] 21. Micro-injection pump combination Detailed Implementation

[0055] In the following description, many technical details are presented to help the reader better understand this application. However, those skilled in the art will understand that the technical solutions claimed in this application can be implemented even without these technical details and various variations, combinations, and modifications based on the following embodiments.

[0056] 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.

[0057] This application relates to a live cell tissue perfusion collection system, the structural diagram of which is shown below. Figure 1-5 As shown, it includes: a fluid flow switching drug delivery module, a constant temperature incubation module, an automatic sample collection module, and a sample low temperature maintenance module 9.

[0058] The fluid switching drug delivery module is configured to switch different liquids at any time according to experimental requirements and pump them evenly into the isothermal incubation module. The isothermal incubation module is configured to provide a closed, constant-temperature incubation fluid environment for live cell tissue perfusion experiments. The automatic sample collection module is configured to receive the liquid samples flowing through the isothermal incubation module and collect the samples into collection well 16 according to a specified time sequence. The sample low-temperature maintenance module 9 maintains the sample collection plate 8 at a relatively low temperature through refrigeration and temperature control to delay sample volatilization and denaturation of the analytes, ensuring sample stability. Automation of operation is achieved through parameter setting and operational control of the entire system. The operation of this system can accurately collect perfusion fluid samples according to the time sequence, improving the accuracy of subsequent detection results and thus obtaining reproducible, high-quality experimental results.

[0059] like Figure 2 and Figure 3 As shown, in an optional embodiment, the fluid flow switching drug delivery module includes two parts: a fluid flow dynamic structure 4 and a fluid flow switching and dispensing structure 3. The fluid flow dynamic structure 4 is configured to provide power for the fluid flow in the live cell tissue perfusion experiment, ensuring the fluid flow within the closed pipe. The fluid flow dynamic structure 4 is located on the fluid flow pipe 1 at the front end of the incubation chamber 6, thereby providing power to the liquid in the dispensing fluid flow pipe 1 to deliver it to the incubation chamber 6. The fluid flow dynamic structure 4 employs an 8-channel peristaltic pump, a diaphragm pump, a micro-injection pump, or an independent multi-channel micro-injection pump combination 21 (such as...). Figure 5 As shown in the figure, the liquid flow switching and dispensing structure 3 adopts a combination of a multi-channel switching valve 11 and a multi-channel dispenser 12. The multi-channel switching valve 11 disperses liquids from different perfusion fluid sources into different incubation chambers 6 through the multi-channel dispenser 12. Figure 3 Figures (1) and (2) show two different structures of the multichannel dispensing device 12, such as T-type or Y-type. The structure of the multichannel dispensing device 12 can also be adjusted according to other experimental needs, as long as it can achieve the function of uniformly dispensing liquid. The liquid flow switching drug delivery module performs closed-loop control according to the specific experimental design requirements, dynamically adjusting the liquid delivery speed, time and timely switching of different liquids to ensure the repeatability and reliability of high-throughput experimental data.

[0060] In an optional embodiment, the constant temperature incubation module consists of eight incubation chambers 6 and a heating temperature control structure. Each incubation chamber 6 is a sealed cavity structure, containing the same number of live cells or ex vivo tissues and solid support material. The perfusion fluid enters from the upper inlet of the incubation chamber 6, perfuses the live cells or ex vivo tissues, and then flows out from the lower outlet of the incubation chamber 6. A screen structure at the outlet ensures that the live cells or ex vivo tissues and solid support material remain inside the incubation chamber 6 throughout the perfusion process. After perfusion, the perfusion sample flows out from the perfusion fluid outlet into the automatic sample collection module. The heating temperature control structure 5 uses a combination of a PTC heating plate and an air bath to ensure that the temperature within the eight incubation chambers 6 is maintained within the set temperature range (room temperature to 45°C).

[0061] In an optional embodiment, the automatic sample collection module consists of a liquid drainage structure and a sample collection time sequence control structure. The liquid drainage structure uses elastic fiber filaments that can be controlled to move linearly as drainage rods 15. The drainage rods 15 make oblique contact and separation with the sample output port 13 and the sidewall of the collection hole, ensuring that the droplets 14 fall into the collection holes 16 of the sample collection plate 8 below, reducing sample liquid spillage outside the collection holes 16 and minimizing the volume error of sample liquid collected in each collection hole 16. The sample collection time sequence control structure consists of a sample collection plate 8, a fixed support plate 10, and a mechanical drive structure. The sample collection plate 8 is fixed on the fixed support plate 10, which can be driven by the mechanical drive structure. The mechanical drive structure consists of a motor and a linear guide rail. During the sampling process, the sample collection plate 8, together with the fixed support plate 10, undergoes horizontal linear displacement and pauses at regular time intervals. This ensures that the collection holes 16 of the sample collection plate 8 are directly opposite the perfusion fluid outlet of the incubation chamber 6 above, facilitating sample collection and ensuring that droplets 14 fall into the collection holes 16 sequentially in chronological order. The speed, movement time, and dwell time of the fixed support plate 10 are determined based on the experimental requirements for the time sequence, the volume of the collection holes 16 on the plate, and the subsequent sample requirements. This ensures that droplets 14 fall accurately into the collection holes 16 in chronological order and enables continuous sampling from multiple collection holes 16, achieving the goal of time-series sampling. The fixed support plate 10 is made of metal and constitutes the metal bath structure component of the sample cryogenic maintenance module 9.

[0062] In an optional embodiment, the sample low-temperature maintenance module 9 consists of a semiconductor cooling structure, a refrigerant circulation structure, and a fixed support plate 10 with metal heat exchange pipes. During operation, the semiconductor cooling structure lowers the temperature of the refrigerant. The refrigerant flows through the metal heat exchange pipes of the fixed support plate to remove heat, and then returns to the semiconductor cooling structure for secondary cooling, achieving unidirectional circulation of the refrigerant. After cooling, the metal fixed support plate 10 further lowers the temperature of the liquid sample in the collection hole 16, thereby realizing the metal bath function, preventing the liquid sample from evaporating and the analyte from denaturing, achieving low-temperature stability of the sample, reducing the impact on experimental results, and improving the reliability of experimental data.

[0063] In an optional embodiment, a PLC industrial control computer may also be included. Through the PLC industrial control computer, users can set and change parameters such as the operation of the peristaltic pump, the operation of the multi-channel switching valve 11, the temperature of the PTC heating plate, the movement of the drainage rod 15, the operation of the drive motor, the position of the collection hole 16 plate, the temperature of the semiconductor refrigeration, and the circulation speed of the refrigerant. The control circuit receives signals from temperature sensors, displacement sensors, and other sensors to control the operation of the peristaltic pump, the multi-channel switching valve 11, the PTC heating plate, the drainage rod 15, the drive motor, the semiconductor refrigeration, and the refrigerant circulation, ultimately realizing the automated operation of the entire system.

[0064] This embodiment provides an efficient, stable, and reproducible live cell tissue perfusion collection system through the above-described method. It is applicable to multiple fields such as drug screening, cell metabolism research, and biomedical experiments, improving experimental efficiency and ensuring the reliability and accuracy of experimental data.

[0065] To better understand the technical solution of this application, another specific embodiment will be described below. The details listed in this example are mainly for ease of understanding and are not intended to limit the scope of protection of this application.

[0066] This embodiment provides a live cell tissue perfusion collection system, which consists of a fluid flow switching drug delivery module, a constant temperature incubation module, and an automatic sample collection and cryopreservation module.

[0067] In this embodiment, the fluid flow switching drug delivery module uses multiple micro-injection pumps as sources of various perfusion fluids 2 and as a fluid flow driving system. Different perfusion fluids are switched via fluid flow pipelines 1 and multi-channel switching valves 11. A multi-stage Y-shaped distributor is used to evenly distribute the perfusion fluid to the incubation chamber 6 of the constant-temperature incubation module. This allows for precise control of fluid flow rate and pressure changes, enabling quantitative control of multiple parameters in perfusion experiments. It further quantifies the conditions for perfusion experiments, improving the accuracy, experimental quality, and reproducibility of perfusion sample collection.

[0068] This embodiment of the constant temperature incubation module consists of a heated metal bath structure and multiple incubation chambers 6, which can be fixed by a bracket. Piping is installed within the metal block to parallelly lay the perfusion fluid input pipes of the incubation chambers 6. Holes are provided as heating tanks for the incubation chambers 6. A PTC ceramic heating plate and a temperature control probe are then configured to form the metal bath heating structure, ensuring that the temperature inside the incubation chambers 6 is maintained within the set temperature range (room temperature to 45°C). The incubation chambers 6 are sealed chambers, each containing the same number of live cells or ex vivo tissues and solid support materials. The perfusion fluid enters from the upper inlet of the incubation chamber 6, perfuses the live cells or ex vivo tissues, and then flows out from the lower outlet of the incubation chamber 6. A screen structure at the outlet ensures that the live cells or ex vivo tissues and solid support materials remain inside the incubation chamber 6 throughout the perfusion process. After perfusion, the perfusion sample flows out from the perfusion fluid outlet into the automatic sample collection module.

[0069] The sample automatic collection and cryopreservation module in this embodiment consists of a liquid drainage structure and a sample cryopreservation structure. For example... Figure 4 As shown in (1)-(3), the liquid drainage structure can be a controllable, elastic drainage rod 15 (which can be a fiber filament or other elastic, slender object). The fiber filament makes oblique contact with and separates from the sample output port and the side wall of the collection hole. Figure 4 (1) Alternatively, a controllable support rod can be used to connect to the drainage plate 17 (which can be a metal sheet or a thin sheet made of other materials) in the form of a spring 18. The drainage plate 17 is in oblique contact with and separates from the sample output port and the side wall of the collection hole. Figure 4 (2) Alternatively, the sample output port 13 is set at an angle, with a section of elastic silicone rubber tube 20 at the front end as a drainage tube. When the sample collection plate 8 moves horizontally and vertically, the end of the drainage tube contacts and separates obliquely from the side wall of the collection hole. Figure 4 (3) In order to ensure that the droplets 14 fall into the collection hole 16 below, avoid the sample liquid from falling outside the collection hole 16, and reduce the error in the volume of sample liquid collected by each collection hole 16.

[0070] The sample cryogenic collection structure consists of a sample collection well plate 8 (e.g., a 96-well plate), a fixed support plate 10, a cryogenic maintenance structure, and a mechanical drive structure. The sample collection well plate 8 is fixed on the fixed support plate 10, which is a metal plate with metal heat exchange pipes inside. The pipes are filled with refrigerant, which can circulate to ensure uniform surface temperature of the fixed support plate 10. Below it is a semiconductor cooling block and its cooling fan. The above structures are integrated into one unit and are driven by the underlying mechanical drive structure to move horizontally and vertically.

[0071] The mechanical drive structure uses a motor and a linear guide rail to perform horizontal linear movement. During the sampling process, the sample collection orifice plate 8, together with other integrated structures, performs horizontal linear displacement and pauses at certain time intervals, so that the collection holes 16 of the sample collection orifice plate 8 are directly opposite the perfusion liquid outlet of the incubation chamber 6 above for sample collection, ensuring that the droplets 14 fall into the collection holes 16 in chronological order. The mechanical drive structure also uses a motor and connecting rod to lift and lower the sample collection orifice plate 8, together with other integrated structures, along an arc. When the collection holes 16 of the sample collection orifice plate 8 are directly opposite the perfusion liquid outlet of the incubation chamber 6 above, the end of the drainage tube can make oblique contact and separation with the side wall of the collection hole, so that the liquid flows down into the collection hole 16 below, avoiding sample liquid from scattering outside the collection hole 16 and reducing the error in the volume of sample liquid collected by each collection hole 16. The speed, movement time, and residence time of the fixed plate 10 will be determined according to the experimental requirements for time series settings, the volume of the collection holes 16 on the collection hole 16 plate, and the required amount of samples for subsequent experiments, so as to ensure that the droplets 14 fall into the collection holes 16 in time sequence and can achieve continuous sampling of multiple collection hole 16 plates to achieve the purpose of sampling according to time series.

[0072] In this embodiment, a PLC industrial control computer may also be included. Through the PLC industrial control computer, users can set and change parameters such as the operation of each micro-injection pump, the operation of the multi-channel switching valve 11, the temperature of the PTC (Positive Temperature Coefficient) heating plate, the operation of the drive motor, the temperature of the semiconductor refrigeration, and the circulation speed of the refrigerant. The PLC industrial control computer receives electrical signals from temperature sensors, displacement sensors, and other sensors to control the operation of structures such as the liquid flow of the micro-injection pump 21, the multi-channel switching valve 11, the PTC heating plate, the drive motor, the position and displacement of the collection hole 16 plate, the semiconductor refrigeration, and the circulation of the refrigerant, ultimately realizing the automated operation of the entire system.

[0073] 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.

[0074] All documents mentioned in this application are considered to be incorporated in their entirety into the disclosure of this application so that they can serve as a basis for modifications 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 live cell tissue perfusion collection system, characterized in that, include: Fluid flow switching drug delivery module, isothermal incubation module, automatic sample collection module, and sample low-temperature maintenance module; The liquid flow switching drug delivery module is configured to switch, divert, and pump different liquids into the constant temperature incubation module at any time according to experimental requirements; The isothermal incubation module is configured to provide a closed, isothermal incubation fluid environment for live cell tissue perfusion experiments; The automatic sample collection module is configured to receive the perfusion fluid sample flowing through the constant temperature incubation module and collect the perfusion fluid sample into the sample collection well plate according to a specified time sequence. The sample low-temperature maintenance module keeps the sample collection well plate at a low temperature to delay the volatilization of the perfusion fluid sample and the denaturation of the analyte therein, thus ensuring the stability of the perfusion fluid sample.

2. The live cell tissue perfusion collection system as described in claim 1, characterized in that, The fluid flow switching drug delivery module comprises two parts: a fluid flow dynamic structure and a fluid flow switching and dispensing structure. The fluid dynamic structure is configured to provide power for the fluid flow in the live cell tissue perfusion experiment, ensuring that the fluid flows within the closed pipe. The liquid flow switching and dispensing structure is configured to automatically switch the type of perfusion fluid input to the constant temperature incubation module according to the experimental design requirements, and then distribute it evenly into the incubation chamber of the constant temperature incubation module via a multi-channel dispenser.

3. The live cell tissue perfusion collection system as described in claim 2, characterized in that, The fluid dynamic structure is configured to quantitatively control the speed, time, and dosage. Depending on the experimental requirements for the quantitative control, a multi-channel or single-channel peristaltic pump, diaphragm pump, micro-injection pump, or independent multi-channel micro-injection pump system is selected to deliver the fluid flow through the pipeline.

4. The live cell tissue perfusion collection system as described in claim 2, characterized in that, The liquid flow switching and dispensing structure is configured to employ one or more multi-channel switching valves, solenoid valves, multi-channel dispensers or combinations thereof, depending on different experimental settings, or to undertake part of the liquid flow switching and dispensing functions by employing a micro-injection pump or a multi-channel micro-injection pump.

5. The live cell tissue perfusion collection system as described in claim 1, characterized in that, The constant temperature incubation module includes multiple incubation chambers and a heating and temperature control structure; The incubation chamber is a closed cavity structure. Each incubation chamber contains the same number of live cells or ex vivo tissues and solid support materials. The upper end of the incubation chamber is provided with a perfusion fluid inlet. The perfusion fluid enters the multiple incubation chambers through the perfusion fluid inlet to perfuse the live cells or ex vivo tissues to provide nutrition or drug treatment. The lower end of the incubation chamber is provided with a perfusion fluid outlet with a screen structure to ensure that the live cells or ex vivo tissues and solid support materials remain in the incubation chamber during the perfusion process. After the perfusion is completed, the perfusion fluid sample flows out from the perfusion fluid outlet to the automatic sample collection module. The perfusion fluid sample includes the metabolic products and secretions of the live cells or ex vivo tissues.

6. The live cell tissue perfusion collection system as described in claim 5, characterized in that, The heating and temperature control structure ensures that the temperature in the multiple incubation chambers remains constant, within the range of room temperature to 45°C. Depending on the different experimental setup requirements, various structures such as air baths, water baths, metal baths, heating rods, and / or temperature sensors are selected to maintain a constant temperature.

7. The live cell tissue perfusion collection system as described in claim 1, characterized in that... The automatic sample collection module includes: a liquid drainage structure and a sample collection time sequence control structure; The liquid drainage structure is configured to ensure that the droplets fall into the collection holes of the sample collection plate below, thereby reducing the amount of sample liquid scattered outside the collection holes and reducing the volume error of the sample liquid collected by each collection hole. The sample collection time series control structure is configured to set the residence time and movement speed of the sample collection well plate according to experimental requirements, so as to collect the sample into the collection well according to the time series requirements and sample collection volume requirements.

8. The live cell tissue perfusion collection system as described in claim 7, characterized in that, The sample collection time sequence control structure is configured such that the collection holes of the sample collection plate are directly opposite the perfusion fluid outlet of the incubation chamber above. The sample collection plate is fixed to a fixed support plate, which is driven by its mechanical drive structure to make horizontal linear displacement and pause at certain time intervals during the sampling process, ensuring that the droplets fall into the collection holes in chronological order. The speed, movement time, and dwell time parameters of the fixed support plate are determined according to the experimental requirements for the time sequence setting, the volume of the collection holes on the collection plate, and the required amount of samples for subsequent experiments.

9. The live cell tissue perfusion collection system as described in claim 7, characterized in that, The liquid drainage structure employs a controllable, elastic drainage rod that makes oblique contact with and separates from the sample output port and the sidewall of the collection hole. Alternatively, a controllable movement support rod can be used to connect to the drainage plate in the form of a spring, wherein the drainage plate makes oblique contact with and separates from the sample output port and the side wall of the collection hole; Alternatively, the sample output port can be angled and have a flexible drainage tube at the front end. By controlling the movement of the drainage tube at the front end of the sample output port and the side wall of the collection hole on the sample collection plate, the end of the drainage tube at the front end of the sample output port can be angled to contact and separate from the side wall of the collection hole. This allows the droplets to fall into the collection hole below, preventing the sample liquid from scattering outside the collection hole and reducing the error in the volume of sample liquid collected in each collection hole.

10. The live cell tissue perfusion collection system as described in claim 1, characterized in that, The sample low-temperature maintenance module is configured to keep the sample collection plate at a relatively low temperature by means of a water bath or metal bath, so as to delay sample volatilization and denaturation of the analyte, and ensure sample stability. The refrigeration principle is achieved by ice-water circulation, semiconductor refrigeration, or semiconductor refrigeration plus refrigerant circulation.

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