Biological organ printing device and printing method
Through the combination of the reagent matrix module, reagent sorting module and printing module of the biological organoid printing device, the automated printing and culture of biological organoids is achieved, solving the problems of uncontrollable operation and environmental pollution in the prior art, and achieving efficient and accurate cell culture and printing.
Patent Information
- Application Number
- CN202510665128.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-22
- Publication Date
- 2025-07-18
AI Technical Summary
The existing bioprinting technology cannot achieve efficient continuous printing of complex models, and the operation process is uncontrollable, resulting in inaccurate experimental results, unable to achieve full integration of printing, culture, growth and imaging, and there is a risk of environmental pollution.
The combination of reagent matrix module, reagent sorting module and printing module is adopted to realize the automated operation of reagents through the liquid extraction needle and power component, combining temperature and CO2 control, ultraviolet sterilization, vision sensors and imaging modules for real-time monitoring, to realize the automated and sterile environment of cell culture.
It realizes accurate and efficient operation of cell culture, reduces artificial errors, eliminates environmental pollution, and completes the full process automation of cell resuspension, printing, culture and imaging, improving operation efficiency and printing accuracy.
Smart Images
Figure CN120330053A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomedical engineering, and particularly relates to a biological organoid printing device and a printing method. Background Art
[0002] Under the control of a computer, bioprinting locates living cells and other biological products, biomaterials, biochemical reagents or biocompatibilities in sequence by layer-by-layer deposition to allow their spatial construction, so as to develop living tissues and organs for tissue engineering, regenerative medicine, and pharmacokinetics, and is more widely used in biological research. A 3D bioprinter is a device that can, under the drive of a digital three-dimensional model, position and assemble biomaterials or cell units according to the principle of additive manufacturing to manufacture products such as medical devices, tissue engineering scaffolds, and tissue organs.
[0003] Cancer has always been the most important factor affecting human health globally. In recent years, there have been more and more researches on targeted new drugs and new tumor immunotherapies related to cancer. However, research with patients as the research object is uncontrollable, and the differences between experimental animals and humans are huge, making it difficult to transfer experimental results. The 2D cell line culture lacks the in-vivo microenvironment and loses some tumor characteristics, resulting in inaccurate results. Currently, these problems have led to extremely low conversion efficiency from basic research to clinical application. The rise of tumor organoids provides a new technical platform for translational medicine. Tumor organoids are organoids cultured from a single tumor sample. This model well preserves tumor heterogeneity and can be amplified, cryopreserved, and genetically modified. Therefore, a large-scale tumor organoid bank has been established currently. Tumor organoid research has become an important tool in tumor basic and clinical research, and is of great significance for revealing the mechanisms of tumor occurrence and development and rapidly evaluating the therapeutic effects of tumor drugs and immune cells.
[0004] However, the current production of tumor organoids and drug screening still remain in the stage of small-scale laboratory research. The production method highly depends on the skills of experimentalists, and the experimental results have large uncontrollability. At the same time, transferring the prepared tumor organoids to 96-well or 384-well plates is time-consuming and laborious, and the operation process is also uncontrollable, easily causing uncontrollable factors such as different sizes of organoids and inconsistent numbers of organoids in each well, resulting in inaccurate experimental results.
[0005] In addition, most of the existing bioprinting technologies print through a single nozzle, and cannot complete the efficient and continuous printing of complex models. When facing operations that require different materials, the nozzle needs to be replaced, which is extremely cumbersome. Moreover, current bioprinters focus on printing, and a fully integrated device that can combine printing, culturing, growing, and imaging has not yet appeared. Therefore, it is necessary to provide an improved biological organoid printing device to solve the above problems. Summary of the Invention
[0006] The object of the present invention is to provide a biological organoid printing device and a printing method. Through the cooperation of a reagent matrix, a reagent sorting module, and a printing module, it is possible to complete the automated operation of cell culture, reduce the error of manual operation, and at the same time prevent the pollution of the cell culture by the environment.
[0007] To achieve the above object, the present invention provides a biological organoid printing device, including:
[0008] A reagent matrix module for storing target formulated reagents and / or target printing reagents;
[0009] A reagent sorting module including a liquid extraction needle and a power assembly. The liquid extraction needle is used to select target formulated reagents and / or target printing reagents from the reagent matrix module; the power assembly is used to control the rotation or vibration of the liquid extraction needle to mix the reagents in the reagent matrix module.
[0010] A printing module connected to the reagent matrix module and / or the liquid extraction needle, for printing the target printing reagent onto the target area of the chip to obtain a biological organoid.
[0011] Further, the reagent sorting module includes a biaxial platform, a gas-liquid dual-use pump, and a connecting pipeline. One end of the gas-liquid dual-use pump is connected to the printing module through the connecting pipeline, and the other end is provided with a liquid extraction needle; the biaxial platform is used to control the movement of the liquid extraction needle to reach different positions of the reagent matrix module, and control the rotation or vibration of the liquid extraction needle to mix the reagents in the reagent matrix module.
[0012] Further, the reagent matrix module includes several groups of reagent bottles for containing reagents to be printed and cleaning liquid; the reagents to be printed are either the already formulated target printing reagents or the unformulated target formulated reagents; when the reagent to be printed is the unformulated target formulated reagent, the reagent sorting module selects the target formulated reagent from the reagent matrix module to formulate the target printing reagent.
[0013] Further, the printing module includes a printing needle, a printing liquid path, and a moving axis; the printing needle is connected to the reagent sorting module or the reagent matrix module through the printing liquid path, for sucking the target printing reagent and then printing;
[0014] The moving axis is used to control the movement of the printing needle in the X-axis, Y-axis, and Z-axis directions.
[0015] Further, the printing module also includes a liquid path selection module and several valves; the liquid path selection module is used to control the opening and closing of the valves to control the printing needle to suck different reagents from the reagent matrix module.
[0016] Further, the biological organ printing device further includes:
[0017] A printing box for accommodating the reagent matrix module, the reagent sorting module, the printing module and the chip;
[0018] A temperature sensor for monitoring the temperature inside the printing box;
[0019] A temperature control element for heating or cooling according to the temperature monitored by the temperature sensor, so that the temperature inside the printing box reaches the target set temperature;
[0020] A CO2 sensing module for monitoring and controlling the CO2 concentration inside the printing box to reach the target concentration.
[0021] Further, the biological organ printing device further includes a vision sensor and an imaging module disposed inside the printing box. The vision sensor is used to track the printing trajectory of the printing module; the imaging module is used to perform cell imaging monitoring on the cell growth status in the chip.
[0022] Further, an ultraviolet sterilization array is also disposed inside the printing box for sterilizing the inside of the printing box.
[0023] The present invention also provides a biological organ printing method, which uses the biological organ printing device described in any one of the above, and includes the following steps:
[0024] S1. Store the target formulated reagent and / or the target printing reagent in the reagent matrix module; when only the target formulated reagent is stored in the reagent matrix module, select the corresponding target formulated reagent through the reagent sorting module to prepare the target printing reagent, and store it in the reagent matrix module;
[0025] S2. The printing module sequentially sucks the target printing reagent from the reagent matrix module and prints it in the target area in the chip. After printing, it is cured to obtain a biological organ.
[0026] Further, it also includes, before the curing, controlling the temperature inside the printing box at 2-8 °C through the temperature control element;
[0027] In step S1, first sterilize the inside of the printing box, and then store the cleaning solution, the target formulated reagent and / or the target printing reagent in the reagent matrix module;
[0028] In step S2, after one kind of target printing reagent is printed, raise the temperature to 37 °C, adjust the carbon dioxide concentration to the target concentration, and then start curing. After curing is completed, detect whether it is qualified through the imaging module. If it is qualified, then repeat step S2 to perform the printing and curing of the next group of target printing reagents;
[0029] Among them, every time the reagent sorting module and the printing module need to replace the reagent, they first clean it with the cleaning solution and then aspirate the next group of reagents.
[0030] Generally speaking, compared with the prior art, the above technical solutions conceived by the present invention mainly have the following technical advantages:
[0031] 1. The bio-organoid printing device provided by the present invention can complete the storage and preparation of target printing reagents through the reagent matrix module and the reagent sorting module, adapt to various printing scenarios, and the printing module can aspirate different reagents from the reagent matrix module to complete the printing of organs with target shapes. The present invention can integrate the preparation, sorting, and printing of reagents, thus accurately and efficiently completing cell culture work, greatly reducing the errors of manual operations, and eliminating the pollution of the environment to cell culture.
[0032] 2. The reagent sorting module provided by the present invention can control the movement of the liquid aspiration needle through a two-axis platform to reach different positions of the reagent matrix module, and can control the rotation or vibration of the liquid aspiration needle to mix the reagents in the reagent matrix module. With such a setting, the present invention can complete the full-process operations of the inoculation and culture of organoids / cells, covering functions such as cell resuspension, printing, culture, culture medium supplementation, and timing imaging, realizing unattended operation and filling the gap that current bio-printers cannot be fully integrated.
[0033] 3. The reagent matrix module of the present invention includes several groups of reagent bottles for storing the reagents to be printed and the cleaning solution, which is convenient for aspirating different reagents as needed. Moreover, when changing to the next group of reagents after each group of reagents is aspirated, it can be cleaned with the cleaning solution, reducing pollution and eliminating the need to frequently change the needle, improving the operation efficiency and convenience. Brief Description of the Drawings
[0034] Figure 1 It is a schematic diagram of the overall structure of the bio-organoid printing device.
[0035] Figure 2 It is a schematic diagram of the partial structure of the bio-organoid printing device.
[0036] Figure 3 It is a schematic diagram of the structure of the reagent matrix and the reagent sorting module.
[0037] Figure 4 It is a schematic diagram of the partial structure of the reagent sorting module and the printing module.
[0038] Figure 5 It is a schematic diagram of the partial structure of the printing module.
[0039] Figure 6 It is a schematic diagram of the structure of the printing module.
[0040] In all the drawings, the same reference numerals are used to denote the same elements or structures, where:
[0041] 1 - Ultraviolet sterilization array; 2 - Reagent matrix module; 21 - Reagent A reagent bottle; 22 - Reagent B reagent bottle; 23 - Reagent C reagent bottle; 24 - Cleaning liquid reagent bottle; 3 - Reagent sorting module; 31 - Biaxial platform; 32 - Gas-liquid dual-purpose pump; 4 - Chip; 5 - Printing module; 51 - Printing needle; 52 - Printing liquid path; 53 - Liquid path selection module; 54 - Moving axis; 55 - Liquid level sensor; 56 - Valve; 6 - Vision sensor; 7 - Imaging module; 8 - Temperature control element; 81 - Temperature sensor; 9 - CO2 sensing module; 10 - Control center; 11 - Data transmission interface; 12 - Power supply. Detailed implementation manners
[0042] In order to make the objectives, technical solutions and advantages of the present invention more clear and understandable, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. In addition, the technical features involved in the various implementation manners of the present invention described below can be combined with each other as long as they do not conflict with each other.
[0043] Please refer to Figures 1-6 as shown, the present invention provides a bio-organ printing device, including:
[0044] A reagent matrix module 2, used for storing target formulated reagents and / or target printing reagents;
[0045] A reagent sorting module 3, including a liquid extraction needle and a power component, the liquid extraction needle is used to select target formulated reagents and / or target printing reagents from the reagent matrix module 2; the power component is used to control the rotation or vibration of the liquid extraction needle to mix the reagents in the reagent matrix module 2;
[0046] A printing module 5, connected to the reagent matrix module 2 and / or the liquid extraction needle, and used for printing the target printing reagent onto a target area of a chip 4 to obtain a bio-organ.
[0047] The printing process of biological organoids mainly includes cell resuspension, cell printing, cell culture, supplementing culture medium, etc. During the printing process, the imaging module 7 automatically focuses and takes pictures of the cell growth status in the chip 4 for live cell imaging to facilitate real-time monitoring of cell growth status. Cell resuspension is achieved through operations such as vibration and pipetting by the reagent sorting module 3 within the reagent matrix module 2 to completely mix the reagents and produce a homogeneous suspension. The reagent raw materials required for cell printing, cell culture, supplementing culture medium, etc. include cells (providing biological activity and tissue regeneration ability), hydrogels (as matrix materials, providing structure and support), and growth factors (promoting cell growth and differentiation and accelerating the tissue regeneration process), which can be formulated into target printing reagents by adding culture medium. After printing, the culture medium can usually be supplemented to further cultivate the printed organ to promote cell proliferation and differentiation.
[0048] Specifically, the biological organoid printing device further includes:
[0049] A printing box for accommodating the reagent matrix module 2, the reagent sorting module 3, the printing module 5, and the chip 4; the purpose is to place each module in a sealed box to facilitate controlling the temperature and CO2 concentration within the box, isolating the external environment to prevent contamination, and improving the cell culture effect.
[0050] A temperature sensor 81 is also provided inside the printing box for monitoring the temperature inside the printing box;
[0051] A temperature control element 8 for heating (37°C, for cell cultivation and solidification) or cooling (2 - 8°C, for reagent storage and printing stages) according to the temperature monitored by the temperature sensor 81 to make the temperature inside the printing box reach the target set temperature;
[0052] A CO2 sensing module for monitoring and controlling the CO2 inside the printing box to reach the target concentration.
[0053] An ultraviolet sterilization array 1 is also provided inside the printing box for sterilizing the inside of the printing box and the chip (such as a graphene chip) to ensure a sterile operating environment. Graphene chip - the printed chip, and cell culture in the chip is completed through the cooperation of the chip and the device.
[0054] Particularly, the biological organoid printing device further includes a vision sensor 6 and an imaging module 7 provided inside the printing box. The vision sensor 6 is used to locate the printing position and track the printing trajectory of the printing module 5 to facilitate improving the printing accuracy; the imaging module 7 is used to automatically focus and take pictures of the cell growth status in the chip for cell imaging monitoring of the cell growth status in the chip to facilitate improving the printing effect.
[0055] Such as Figure 3As shown, the reagent matrix module 2 includes several groups of reagent bottles for containing the reagents to be printed and the cleaning liquid; the reagents to be printed are the target printing reagents that have been prepared or the target preparation reagents that have not been prepared; when the reagent to be printed is the target preparation reagent that has not been prepared, the reagent sorting module 3 selects the target preparation reagent from the reagent matrix module 2 to prepare the target printing reagent.
[0056] Specifically, the reagent matrix module 2 is respectively fixed on two side walls (preferably two opposite side walls) inside the printing box, as Figure 1 and 3 shown, including reagent A reagent bottle 21, reagent B reagent bottle 22, reagent C reagent bottle 23, cleaning liquid reagent bottle 24, etc., which are respectively fixed on the side walls for containing different reagents.
[0057] The reagent sorting module 3 is arranged above the reagent matrix module 2 for reagent selection, piercing and liquid transmission. Specifically, it includes a biaxial platform 31, a gas-liquid dual-purpose pump 32 and a connecting pipeline. One end of the gas-liquid dual-purpose pump 32 is connected to the printing module 5 through the connecting pipeline, and the other end is provided with a liquid extraction needle; the biaxial platform 31 is used to control the movement of the liquid extraction needle to reach different positions of the reagent matrix module 2, and control the rotation or vibration of the liquid extraction needle to mix the reagents in the reagent matrix module 2. For example, the biaxial platform 31 controls the liquid extraction needle to move horizontally above the target reagent bottle, and then moves downward to pierce the protective film on the reagent bottle until the liquid extraction needle reaches the bottom of the reagent bottle (without direct contact), and then controls the liquid extraction needle to rotate or vibrate (for 10 - 30 s) to make the reagents in the reagent bottle mixed evenly (such as cell suspension, etc., when refrigerated, the cell suspension will have some precipitation).
[0058] As Figures 5-6 shown, the printing module 5 includes a printing needle 51, a printing liquid path 52 and a moving shaft 54; the printing needle 51 is connected to the reagent sorting module 3 or the reagent matrix module 2 through the printing liquid path 52 for sucking the target printing reagent and printing in the chip 4 (it can directly suck from the reagent matrix module 2 or through the reagent sorting module 3). The printing needle 51 and the printing liquid path 52 are quick-release modules to reduce the contamination of multiple reuses.
[0059] The moving axis 54 is used to control the movement of the printing needle 51 in the X-axis, Y-axis, and Z-axis directions for printing the target area and shape. The printing module 5 further includes a liquid path selection module 53 and a plurality of valves 56; the liquid path selection module 53 is used to control the opening and closing of the valves 56 to control the printing needle 51 to suck different reagents from the reagent matrix module 2. The liquid path selection module 53 can realize the selection of liquids and the liquid control of multiple valves 56. Through the control of several valves 56, functions such as single-pass, multi-channel, and cleaning are completed.
[0060] Specifically, a power supply and a switch are provided on the housing of the printing box for controlling the energy of the device. A control center display screen and a data transmission port are provided on the outer shell. The control center display screen is used to set the basic parameters of temperature and CO2, and can also be used to monitor the signals returned by the vision sensor in real time. The data transmission port is connected to a computer for transmitting the operating status of the device and the data of the imaging module 7 to assist the computer to complete the unattended fully automated printing process.
[0061] The present invention also provides a method for printing biological organoids, using the biological organoid printing device described in any one of the above, including the following steps:
[0062] S1. Place the target formulated reagent and / or target printing reagent in the reagent matrix module 2; when only the target formulated reagent is stored in the reagent matrix module 2, select the corresponding target formulated reagent through the reagent sorting module 3 to formulate the target printing reagent and store it in the reagent matrix module 2;
[0063] S2. The printing module 5 sequentially sucks the target printing reagent from the reagent matrix module 2 and prints it in the target area in the chip 4, and after printing is completed, it is cured to obtain biological organoids.
[0064] Further, before the curing, the temperature in the printing box is controlled at 2-8 °C by the temperature control element 8;
[0065] In step S1, first sterilize the inside of the printing box, and then place the cleaning solution, target formulated reagent, and / or target printing reagent in the reagent matrix module 2;
[0066] In step S2, after one kind of target printing reagent is printed, the temperature is raised to 37 °C, the carbon dioxide concentration is adjusted to the target concentration, and then curing starts. After curing is completed, the imaging module 7 is used to detect whether it is qualified. If it is qualified, then step S2 is repeated to perform the printing and curing of the next group of target printing reagents;
[0067] Among them, each time the reagent sorting module 3 and the printing module 5 need to replace the reagent, they are first cleaned with the cleaning solution and then the next group of reagents is sucked.
[0068] The printing method of the present invention will be elaborated in detail through specific embodiments below.
[0069] Example 1 (Reagent with configured concentration)
[0070] A method for printing biological organoids includes the following steps:
[0071] 1. Select the mode with the reagent configured on the biological organoid printing device. According to the preset temperature (2 - 8°C), the temperature control element 8 starts to work. Wait for the temperature sensor 81 to detect that the set temperature is reached, and the control center prompts that it is ready.
[0072] After the temperature control element 8 starts to cool down, the ultraviolet sterilization array 1 (LED) also starts to be activated for sterilization for 15 - 30 minutes.
[0073] 2. Place the chip 4 into the biological organoid printing device, push the reagent matrix module 2 into the device. After locking the buckle, select several types of cell liquids to be printed preset in the control center, set the volume and speed (here only demonstrate the implementation method of printing two types of cells) and the area and shape of the chip 4 to be printed, and then click start.
[0074] 3. The motor drives the module with the needle (liquid extraction needle) to move above the set reagent. The motor drives the module to move downward, puncture the protective film above the reagent, and the needle reaches the bottom of the reagent bottle (without direct contact).
[0075] 4. Continuously blow for a period of time (10 - 30 s) to completely mix the reagent, generating a homogeneous suspension (such as cell suspension, etc. When refrigerated, the cell suspension will have some precipitation).
[0076] 5. Driven by the moving platform, the printing needle moves to the area of the chip 4 preset by the device, and the vision sensor 6 returns a signal to prompt whether it has reached the ready position.
[0077] 6. Through continuous correction by the vision sensor 6, complete the printing of the path (dot matrix array or path, etc.) in the specified area.
[0078] 7. After completing the printing of one type of cell liquid, the preset program will drive the motor to draw out the needle from the first reagent bottle and return to the initial position, and then move to the reagent bottle containing the cleaning liquid (which can remove cell or organoid residues). Repeat step (3), and the motor rotates forward and backward several times to fully clean the needle.
[0079] 8. Driven by the motor, move to the second printing liquid. The motor drives the module with the needle to move above the set reagent. The motor drives the module to move downward, puncture the protective film above the reagent, and after the needle reaches the bottom of the reagent bottle (without direct contact), repeat steps 4, 5, and 6.
[0080] 9. Start with heating and carbon dioxide, rise to 37 °C within 15 minutes, the sensor receives the signal and starts to enter the curing stage (30 minutes).
[0081] 10. After curing is completed, the imaging module 7 detects whether there is an obvious three-dimensional structure. After the detection is completed, repeat steps 3, 4, 5, 6, and 7. The culture medium is added in this process, and the culture medium can be set to be replenished regularly to achieve automatic liquid replenishment.
[0082] 11. Connect to the computer, store and transmit data in real time, monitor the camera, and complete unattended and remote monitoring.
[0083] Example 2 (reagents with unconfigured concentration)
[0084] A method for printing biological organoids includes the following steps:
[0085] 1. Set the mode of unconfigured reagents on the printing device, set the total volume of the reagents, set the main reagent (reagent A) and the diluent (reagent B), and the storage reagent bottle (reagent C) for the mixed liquid. (Of course, the quantity of this reagent can be expanded and increased according to the actual situation), and the default last one is the cleaning solution;
[0086] The equipment starts to work according to the preset temperature (2 - 8 °C), and the temperature control element 8 waits for the temperature sensor 81 to detect the set temperature, and the control center prompts readiness.
[0087] After the temperature module starts to cool down, the LED ultraviolet sterilization array starts to start for 15 - 30 minutes of sterilization.
[0088] 2. Put the chip 4 into the equipment, push the reagent matrix module 2 into the equipment, after the buckle is locked, select several kinds of cell liquids to be printed preset in the control center, and the set volume and speed (here only demonstrate the mixing and spotting culture process of one kind of cell on the chip, adding cell types, the method is similar), and the area and shape to be printed on the chip 4, and then click start.
[0089] 3. The motor drives the module with the needle to move above the set reagent A reagent bottle 21, the motor drives the module to move downward, pierces the protective film above the reagent bottle, the needle reaches the bottom of the reagent bottle (without direct contact), and the equipment extracts the calculated volume of reagent A.
[0090] 4. The motor drives the module with the needle to move above the set reagent C reagent bottle 23, the motor drives the module to move downward, pierces the protective film above the reagent, the needle reaches the bottom of the reagent bottle (without direct contact), and the equipment puts the extracted calculated reagent A into reagent bottle C.
[0091] 5. The motor drives the module with the needle to move above the cleaning liquid reagent bottle 24 (a reagent bottle that can remove cell or organoid residues). The motor drives the module to move downward, puncture the protective film above the reagent, and the needle reaches the bottom of the reagent bottle (without direct contact). The motor rotates forward and backward several times to fully clean the needle and completely drain the waste liquid.
[0092] 6. Repeat steps 3 and 4 to move Reagent B into Reagent C, mix them, and continuously pipette for a period of time (10 - 30 s) to fully mix the reagents and produce a homogeneous suspension (such as a cell suspension. When refrigerated, there will be some precipitation in the cell suspension). At this time, the liquid mixing is completed.
[0093] 7. Perform printing. The printing needle, driven by the moving platform, moves to the preset chip 4 area of the device. The vision sensor returns a signal to indicate whether it has reached the ready position. Through continuous correction by the vision sensor 6, the printing of the specified area (dot matrix array or path, etc.) is completed.
[0094] 7. After completing the printing of one type of cell solution, the preset program will drive the motor to withdraw the needle from the first reagent bottle and return it to the initial position, and then move to the reagent bottle containing the cleaning liquid (which can remove cell or organoid residues). The motor rotates forward and backward several times to fully clean it;
[0095] 8. Driven by the motor, move to the second printing liquid. The motor drives the module with the needle to move above the set reagent. The motor drives the module to move downward, puncture the protective film above the reagent, and after the needle reaches the bottom of the reagent bottle (without direct contact), repeat steps 3, 4, 5, 6, and 7 (to complete the printing of multiple cells or organoids).
[0096] 9. Start heating and supply carbon dioxide. Raise the temperature to 37 °C within 15 min. When the sensor receives the signal, start entering the curing stage (30 min).
[0097] 10. After curing is completed, the imaging module 7 detects whether there is an obvious three-dimensional structure. After the detection is completed, repeat steps 3, 4, 5, 6, and 7. In this process, the culture medium is added, and the timing of supplementing the culture medium can be set to achieve automatic liquid replenishment.
[0098] 11. Connect to the computer, store and transmit data in real time, and monitor the camera to complete unattended and remote monitoring.
[0099] It is easy for those skilled in the art to understand that the above are only preferred embodiments of the present invention and are not used to limit the present invention. Any modifications, equivalent replacements, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A biological organoid printing device, characterized in that, Comprising: A reagent matrix module (2) for storing target formulated reagents and / or target printing reagents; A reagent sorting module (3), including a liquid extraction needle and a power component, the liquid extraction needle being used to select target formulated reagents and / or target printing reagents from the reagent matrix module (2); the power component being used to control the rotation or vibration of the liquid extraction needle to mix the reagents in the reagent matrix module (2); A printing module (5), connected to the reagent matrix module (2) and / or the liquid extraction needle, for printing the target printing reagent onto the target area of the chip (4) to obtain bio-organoids.
2. The bio-organoid printing device according to claim 1, wherein The reagent sorting module (3) includes a biaxial platform (31), a gas-liquid dual-use pump (32) and a connecting pipeline; one end of the gas-liquid dual-use pump (32) is connected to the printing module (5) through the connecting pipeline, and the other end is connected to the liquid extraction needle; the biaxial platform (31) is used to control the movement of the liquid extraction needle to reach different positions of the reagent matrix module (2).
3. The bio-organoid printing device according to claim 2, characterized in that, The reagent matrix module (2) includes several groups of reagent bottles for containing reagents to be printed and cleaning liquid; the reagents to be printed are either the already formulated target printing reagents or the unformulated target formulated reagents; when the reagent to be printed is the unformulated target formulated reagent, the reagent sorting module (3) selects the target formulated reagent from the reagent matrix module (2) to prepare the target printing reagent.
4. The bio-organ printing device according to claim 1, characterized in that, The printing module (5) includes a printing needle (51), a printing liquid path (52) and a moving shaft (54); the printing needle (51) is connected to the reagent sorting module (3) or the reagent matrix module (2) through the printing liquid path (52) for sucking the target printing reagent and then printing; The moving shaft (54) is used to control the movement of the printing needle (51) in the X-axis, Y-axis and Z-axis directions.
5. The bio-organoid printing device according to claim 4, characterized in that, The printing module (5) further includes a liquid path selection module (53) and several valves (56); the liquid path selection module (53) is used to control the opening and closing of the valves (56) to control the printing needle (51) to suck different reagents from the reagent matrix module (2).
6. The bio-organ printing device according to any one of claims 1-5, characterized in that, The bio-organoid printing device further includes: A printing box for accommodating the reagent matrix module (2), the reagent sorting module (3), the printing module (5) and the chip (4); A temperature sensor (81) for monitoring the temperature inside the printing box; A temperature control element (8) for heating or cooling according to the temperature monitored by the temperature sensor (81) so that the temperature inside the printing box reaches the target set temperature; A CO2 sensing module for monitoring and controlling the CO2 inside the printing box to reach the target concentration.
7. The bio-organoid printing device according to claim 6, wherein, The bio-organoid printing device further includes a vision sensor (6) and an imaging module (7) arranged inside the printing box, the vision sensor (6) being used to track the printing trajectory of the printing module (5); the imaging module (7) being used to perform cell imaging monitoring on the cell growth status inside the chip.
8. The bio-organoid printing device according to claim 6, wherein An ultraviolet sterilization array (1) is further provided in the printing box body for sterilizing the interior of the printing box body.
9. A method for printing biological organoids, characterized in that, Using the bio-organ printing device according to any one of claims 1-8, comprising the following steps: S1. Store the target preparation reagent and / or the target printing reagent in the reagent matrix module (2); when only the target preparation reagent is stored in the reagent matrix module (2), select the corresponding target preparation reagent through the reagent sorting module (3) to prepare the target printing reagent, and store it in the reagent matrix module (2); S2. The printing module (5) sequentially sucks the target printing reagent from the reagent matrix module (2) and prints it in the target area in the chip (4), and after printing is completed, it is cured to obtain a bio-organ.
10. The method for printing biological organoids according to claim 9, characterized in that, It further includes that before the curing, the temperature in the printing box body is controlled at 2-8 °C through the temperature control element (8); In step S1, first sterilize the interior of the printing box body, and then store the cleaning solution, the target preparation reagent and / or the target printing reagent in the reagent matrix module (2); In step S2, after one kind of target printing reagent is printed, raise the temperature to 37 °C, adjust the carbon dioxide concentration to the target concentration, and then start curing. After curing is completed, detect whether it is qualified through the imaging module (7). If it is qualified, then repeat step S2 to perform the printing and curing of the next group of target printing reagents; Among them, each time the reagent sorting module (3) and the printing module (5) need to replace the reagent, they are first cleaned with the cleaning solution and then the next group of reagents is sucked.