Fully-automatic high-throughput blood testing system
Patent Information
- Application Number
- PCT/CN2025/086221
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2025-03-24
- Filing Date
- 2025-03-31
- Publication Date
- 2026-10-01
Smart Images
Figure CN2025086221_01102026_PF_FP_ABST
Abstract
Description
A fully automated high-throughput blood testing system Technical Field
[0001] This invention relates to the field of blood testing system technology, and in particular to a fully automated high-throughput blood testing system. Background Technology
[0002] Currently, blood sample processing, especially component partitioning and preparation for flow cytometry analysis, largely relies on manual operations. The separation of the leukocyte layer and the preparation of flow cytometry samples are particularly time-consuming. Specifically, during blood centrifugation, a thin fibrin membrane forms in the centrifuge tube. A sterile pipette tip is needed to carefully separate the upper plasma layer before aspirating it, avoiding penetration of the membrane and contact with the underlying blood cells. Furthermore, preparing flow cytometry samples involves multiple steps such as repeated centrifugation, mixing, and cell resuspending. The efficiency of these operations is closely related to the operator's skill level, highlighting the high level of manual skill required. Manual blood component separation and flow cytometry analysis experiments have limitations in terms of pipetting accuracy, efficiency, reproducibility, and safety risks. Summary of the Invention
[0003] The purpose of this invention is to provide a fully automated high-throughput blood testing system to solve the problems existing in the prior art and improve testing efficiency and the accuracy of test results.
[0004] To achieve the above objectives, the present invention provides the following solution:
[0005] This invention provides a fully automated high-throughput blood testing system, comprising: a chassis, a feed drawer, a first centrifuge, a transfer mechanism, a pipetting workstation, a flow cytometry pretreatment mechanism, a flow cytometer, and an industrial control computer. The feed drawer is equipped with a material rack for holding blood collection tubes containing whole blood. The feed drawer is slidably mounted on the chassis to feed or remove the blood collection tubes from the chassis. The first centrifuge is located inside the chassis for centrifuging the blood collection tubes containing whole blood. The transfer mechanism is located inside the chassis and can transfer the blood collection tubes from the feed drawer to the first centrifuge. The transfer mechanism can also transfer the centrifuged blood collection tubes to the pipetting workstation. The pipetting workstation is located inside the chassis and is equipped with a perforated plate. The pipetting workstation is used to scan and open the transferred blood collection tubes. The system includes: capping blood collection tubes; identifying the location of the leukocyte layer in each blood collection tube; aspirating the leukocyte layer and adding it to a multi-well plate; and capping the blood collection tubes. A flow cytometry pretreatment mechanism is located within the chassis. This mechanism is used to add reagents, centrifuge, shake, and incubate the multi-well plate containing the leukocyte layer at a controlled temperature. A flow cytometer is located within the chassis. The transport mechanism is also used to transport the pretreated multi-well plate to the flow cytometer, which performs flow cytometry analysis and outputs the results. The industrial control computer controls the operation of the first centrifuge, the transport mechanism, the pipetting workstation, the flow cytometry pretreatment mechanism, and the flow cytometer.
[0006] Preferably, the feed drawer can accommodate 12 8-hole material racks; the transfer mechanism includes material rack grippers, blood collection tube grippers, a material rack gripper drive mechanism, and a blood collection tube gripper drive mechanism. The material rack gripper drive mechanism can drive the material rack grippers to transfer the material racks in the feed drawer to the centrifuge transfer position. The blood collection tube gripper drive mechanism can drive the blood collection tube grippers to clamp the blood collection tubes in the material racks on the centrifuge transfer position into the first centrifuge and to clamp the centrifuged blood collection tubes out of the first centrifuge and put them back into the material racks on the centrifuge transfer position.
[0007] The transfer mechanism also includes a pipetting workstation rack plate and a rack plate driving mechanism. The rack grippers can transport the rack carrying the centrifuged blood collection tubes to the pipetting workstation rack plate, and the rack plate driving mechanism drives the pipetting workstation rack plate carrying the rack to move into the pipetting workstation.
[0008] Preferably, the pipetting workstation includes workstation grippers, a barcode scanner, a workstation gripper drive mechanism, a blood collection tube fixing clamp, a pipette, a pipette drive device, a multi-well plate placement position, and a recognition camera; a multi-well plate is placed at the multi-well plate placement position; the blood collection tube fixing clamp can clamp and release the body of the blood collection tube; the recognition camera is located on one side of the blood collection tube fixing clamp; the recognition camera is used to capture image information of the blood collection tube on the blood collection tube fixing clamp and transmit it to the industrial control computer; the industrial control computer determines the position information of the white blood cell layer inside the blood collection tube based on the image information of the blood collection tube; the workstation grippers can clamp and release the body and cap of the blood collection tube on the material rack plate of the pipetting workstation; the workstation gripper drive mechanism can... The workstation gripper is driven to move, and the range of motion of the workstation gripper covers the pipetting workstation material rack plate and the blood collection tube fixing clamp. The workstation gripper driving mechanism can also drive the workstation gripper to rotate. The pipette driving device can drive the pipette to move, and the range of motion of the pipette covers the multi-well plate placement position and the blood collection tube fixing clamp. The industrial control computer controls the pipette driving device to move the inlet and outlet of the pipette to the white blood cell layer according to the position information of the white blood cell layer in the blood collection tube to aspirate the white blood cell layer. The barcode scanner is integrated with the workstation gripper and located on one side of the workstation gripper. The barcode scanner is used to scan the identification code on the blood collection tube.
[0009] Preferably, multiple workstation grippers, barcode scanners, blood collection tube fixing clamps, and pipettes are provided. Multiple workstation grippers are integrated side-by-side to form a multi-channel gripper; multiple barcode scanners are integrated side-by-side to form a multi-channel barcode scanner; multiple blood collection tube fixing clamps are integrated side-by-side to form a multi-channel fixing clamp; multiple pipettes are integrated side-by-side to form a multi-channel pipette; one workstation gripper corresponds to one barcode scanner and one blood collection tube fixing clamp; one pipette corresponds to one blood collection tube fixing clamp.
[0010] Preferably, the flow cytometry pretreatment mechanism includes a reagent storage mechanism, a temperature control module, an oscillation module, a second centrifuge, multi-well plate grippers, and a multi-well plate gripper driving mechanism. The reagent storage mechanism is used to store the reagents required for pretreatment. The temperature control module is used to perform temperature-controlled incubation of the samples in the multi-well plate. The oscillation module is used to oscillate the samples in the multi-well plate. The second centrifuge is used to centrifuge the samples in the multi-well plate. The stroke of the pipette also covers the reagent storage mechanism. The multi-well plate grippers can clamp and release the multi-well plate. The multi-well plate gripper driving mechanism drives the multi-well plate grippers to move, so that the stroke of the multi-well plate grippers covers the multi-well plate placement position, the temperature control module, the oscillation module, and the second centrifuge.
[0011] Preferably, a plurality of pipettes are arranged at equal intervals along the horizontal direction, each pipette is vertically arranged, the inlet and outlet of the pipette are constructed at the bottom of the pipette, and the pipette driving device is capable of individually driving each of the multi-channel pipettes to move up and down, and is capable of changing the horizontal position of all the pipettes in the multi-channel pipette to achieve synchronous adjustment of the spacing between any two adjacent pipettes.
[0012] Preferably, the transfer mechanism further includes a well plate support frame and a well plate support frame drive mechanism. The well plate support frame is used to carry the well plate. The stroke of the well plate gripper covers the well plate support frame. The well plate support frame drive mechanism can drive the well plate support frame to move within the range of motion of the material rack gripper. The material rack gripper drive mechanism can drive the material rack gripper to clamp the well plate and send the well plate into the flow cytometer.
[0013] Preferably, the chassis is equipped with an ultraviolet disinfection module, and part of the chassis wall is constructed as a viewing window, which is made of ultraviolet-resistant organic glass; the chassis is also equipped with an H14 grade negative pressure HEPA filter module.
[0014] Preferably, the first centrifuge and the second centrifuge are housed inside the casing and positioned on the ground by a positioning frame independent of the casing.
[0015] Preferably, the chassis is divided into a first outer shell and a second outer shell, both of which have an open structure on opposite sides. The open structures of the first and second outer shells are joined together to form the chassis. A central support plate is fixedly installed in the middle of both the first and second outer shells. The first centrifuge and the flow cytometer are disposed inside the first outer shell and located below the central support plate. The feed rack gripper, blood collection tube gripper, feed rack gripper drive mechanism, blood collection tube gripper drive mechanism, centrifuge transfer station, and feed drawer are all disposed inside the first outer shell and located above the central support plate. The pipetting workstation and the flow cytometry pretreatment mechanism are all disposed inside the second outer shell and located above the central support plate. The second centrifuge is disposed inside the second outer shell and located below the central support plate.
[0016] The present invention achieves the following technical effects compared to the prior art:
[0017] The detection system provided by this invention is fully automated, requires no manual intervention, can continuously process a large number of samples, significantly shortens the experimental cycle, and is suitable for large-scale screening and diagnosis.
[0018] This invention automates the execution of standard operating procedures, ensuring consistency of experimental conditions each time and improving the reproducibility of results.
[0019] This invention employs precision mechanical and optical components to automatically identify blood layers, precisely control dispensing volume and reagent addition, reduce human error, and improve the accuracy of experimental results.
[0020] This invention's closed-loop automated operation reduces the opportunity for operators to directly contact biological samples and harmful reagents, thereby lowering the risk of laboratory safety accidents.
[0021] This invention automatically records experimental data, facilitating tracking, management, and analysis, and accelerating report generation. Attached Figure Description
[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0023] Figure 1 is a schematic diagram of the structure of the fully automated high-throughput blood testing system provided in an embodiment of the present invention;
[0024] Figure 2 is the right view of Figure 1;
[0025] Figure 3 is a view of the first centrifuge after the front cover plate of the first centrifuge in Figure 2 has been removed;
[0026] Figure 4 is a front view of the second centrifuge after the front cover plate has been removed from Figure 1;
[0027] Figure 5 is a left view of Figure 1 after removing the cover plate on one side of the flow cytometer;
[0028] Figure 6 is a schematic diagram of the internal structure of the chassis in Figure 1;
[0029] Figure 7 is a schematic diagram of the internal structure of the first outer shell;
[0030] Figure 8 is a partial structural diagram of the transfer mechanism;
[0031] Figure 9 is a schematic diagram of the internal structure of the second outer shell;
[0032] Figure 10 is a top view of the internal structure of the second outer shell;
[0033] Figure 11 is a bottom view of the internal structure of the chassis;
[0034] In the diagram: 1-First outer casing; 2-Second outer casing; 3-Industrial control computer; 4-Feed drawer; 5-First centrifuge; 6-Pipeline workstation window; 7-Solid waste box door; 8-Second centrifuge; 9-Electrical control box door; 10-Flow cytometer; 11-Transfer mechanism; 12-H14 grade negative pressure HEPA filter module; 13-Pickler gripper drive mechanism; 14-Collection tube gripper drive mechanism; 15-Secondary drawer structure; 16-Positioning frame; 17-Pickler gripper; 18-Collection tube gripper; 19-Workstation gripper; 20-Workstation gripper drive mechanism; 21-Collection tube fixing clamp; 22-Pipette; 23-Pipette drive device; 24-Polymer plate; 25-Oscillation module; 26-Temperature control module; 27-Pipeline workstation picker plate; 28-Polymer plate gripper; 29-Polymer plate gripper drive mechanism; 30-Pickler plate drive mechanism; 31-Chassis. Detailed Implementation
[0035] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0036] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.
[0037] The embodiments of the present invention are described below with reference to Figures 1 to 11.
[0038] This invention provides a fully automated high-throughput blood testing system for fully automated processing and flow cytometry analysis of blood samples in the field of early cancer screening and diagnosis. The system includes: a chassis 31, a feed drawer 4, a first centrifuge 5, a transfer mechanism 11, a pipetting workstation, a flow cytometry pretreatment mechanism, a flow cytometer 10, and an industrial control computer 3. The feed drawer 4 is equipped with a material rack, which serves as a carrier for holding blood collection tubes containing whole blood. The feed drawer 4 is slidably mounted on the housing 31 to feed the blood collection tubes into or remove them from the housing 31. The housing 31 has a drawer opening for feeding, and the feed drawer 4 is slidably mounted on the housing 31 via a linear guide rail. Sliding the feed drawer 4 inwards into the housing 31 allows the material rack to be fed into the housing 31, while pulling the feed drawer 4 outwards allows the material rack to be removed from the drawer opening. The first centrifuge 5 is located inside the housing 31 and is used to centrifuge the blood collection tubes containing whole blood to separate the whole blood into layers. The transfer mechanism 11 is located inside the housing 31 and can transfer the blood collection tubes from the feed drawer 4 to the first centrifuge 5. The transfer mechanism 11 can also transfer the centrifuged blood collection tubes to the pipetting workstation. The pipetting workstation is located inside the housing 31 and is used for pipetting operations. The station is equipped with a multi-well plate 24. The pipetting workstation is used to scan the barcode of the transported blood collection tubes, open the caps of the blood collection tubes, identify the position of the white blood cell layer in each blood collection tube, aspirate the white blood cell layer and add it to the multi-well plate 24, and close the caps of the blood collection tubes. The flow cytometry pretreatment mechanism is located in the chassis 31. The flow cytometry pretreatment mechanism is used to add reagents, centrifuge, shake and incubate the multi-well plate 24 with the white blood cell layer. The flow cytometer 10 is located in the chassis 31. The transport mechanism 11 is also used to transport the pretreated multi-well plate 24 to the flow cytometer 10. The flow cytometer 10 performs flow cytometry analysis and outputs the analysis results. The industrial control computer 3 controls the operation of the first centrifuge 5, the transport mechanism 11, the pipetting workstation, the flow cytometry pretreatment mechanism and the flow cytometer 10, and receives information from each module, such as the barcode information in the pipetting workstation and the flow cytometry analysis results information of the flow cytometer 10.
[0039] The detection system provided by this invention only requires placing the blood collection tube containing the sample to be processed onto the rack in the feed drawer 4 and pushing it into the chassis 31. Subsequent processes such as whole blood centrifugation, blood collection tube transfer, pipetting, pretreatment, delivery to the flow cytometer 10, and flow cytometry analysis are all automatically performed by the corresponding modules controlled by the industrial control computer 3. This improves experimental efficiency. Furthermore, the automated execution of standard operating procedures ensures consistency of experimental conditions for each experiment, improving the reproducibility of results. This invention employs precision mechanical and optical components to automatically identify blood stratification, accurately control dispensing volume and reagent addition, reduce human error, and improve the accuracy of experimental results. The closed-loop automated operation of this invention reduces the opportunity for operators to directly contact biological samples and harmful reagents, lowering the risk of laboratory safety accidents.
[0040] Note: The blood collection tube consists of a tube body and a cap. After sampling, the cap is screwed on and fixed to the top of the tube body. That is, when the blood collection tube is placed in the feed drawer 4, it is in the sealed state (the cap is screwed on and fixed to the top of the tube body).
[0041] In some embodiments, the feed drawer 4 can accommodate 12 8-hole material racks; the transfer mechanism 11 includes a material rack gripper 17, a blood collection tube gripper 18, a material rack gripper drive mechanism 13, and a blood collection tube gripper drive mechanism 14. The material rack gripper drive mechanism 13 can drive the material rack gripper 17 to transfer the material rack in the feed drawer 4 to the centrifuge transfer position. The blood collection tube gripper drive mechanism 14 can drive the blood collection tube gripper 18 to clamp the blood collection tubes in the material rack on the centrifuge transfer position into the first centrifuge 5 and to clamp the centrifuged blood collection tubes out of the first centrifuge 5 and put them back into the material rack on the centrifuge transfer position. The transfer mechanism 11 also includes a pipetting workstation material rack plate 27 and a material rack plate drive mechanism 30. The material rack gripper 17 can transport the material rack carrying the centrifuged blood collection tubes to the pipetting workstation material rack plate 27. The material rack plate drive mechanism 30 drives the pipetting workstation material rack plate 27 carrying the material rack to move into the pipetting workstation.
[0042] This embodiment uses an 8-well rack as the carrier, allowing for the feeding of 96 samples at a time, achieving high-throughput detection. In some examples, the first centrifuge 5 can centrifuge a maximum of 32 blood collection tubes at a time. Therefore, in this embodiment, the rack grippers 17 transport 4 racks (32 blood collection tubes) to the centrifuge transfer position each time. The blood collection tube grippers 18 feed the blood collection tubes (with caps) into the first centrifuge 5 one by one. After centrifugation, the blood collection tube grippers 18 automatically remove the blood collection tubes one by one and return them to the rack. If there are fewer than 32 blood collection tubes, the first centrifuge 5 automatically calculates the centrifuge rotor balance; therefore, the model of the first centrifuge 5 needs to have an automatic balance function.
[0043] It is understood that in the above embodiments, the material rack gripper drive mechanism 13 and the blood collection tube gripper drive mechanism 14 are both preferably three-dimensional moving mechanisms. The three-dimensional moving mechanism can drive the material rack gripper 17 and the blood collection tube gripper 18 to move in three-dimensional space. Specifically, the motion principle of the three axes in the three-dimensional moving mechanism can be ball screw, synchronous belt drive, gear and rack drive, etc.
[0044] For ease of understanding, the three directions of movement of the three-dimensional moving mechanism are defined as X, Y and Z.
[0045] In one example, both the material rack gripper drive mechanism 13 and the blood collection tube gripper drive mechanism 14 include an X-axis drive mechanism, a Z-axis drive mechanism, and a Y-axis drive mechanism. The Y-axis drive mechanism is located at the free end of the X-axis drive mechanism, and the Z-axis drive mechanism is located at the free end of the Y-axis drive mechanism. The free end of the Z-axis drive mechanism is connected to the blood collection tube gripper 18 or the material rack gripper 17. Alternatively, the Z-axis drive mechanism can be located at the free end of the X-axis drive mechanism, and the Y-axis drive mechanism can be located at the free end of the Z-axis drive mechanism. Specifically, the X-axis drive mechanism, Z-axis drive mechanism, and Y-axis drive mechanism can use motors and transmission mechanisms to drive their respective free ends to move along their respective axes. The transmission mechanism can be a ball screw, synchronous belt drive, rack and pinion drive, etc.
[0046] More specifically, taking the blood collection tube clamp drive mechanism 14 as an example, the free end travel of the X-axis drive mechanism is 1100mm, the transmission mechanism is a ball screw, and the power motor is a 400W servo motor; the free end travel of the Y-axis drive mechanism is 400mm, the transmission mechanism is a synchronous belt, and the power motor is a 57 closed-loop stepper motor; the free end travel of the Z-axis drive mechanism is 380mm, the transmission mechanism is a helical gear rack and double-speed belt mechanism, and the power motor is a 57 closed-loop stepper motor with brake. The three-dimensional moving mechanism provided in this embodiment allows the blood collection tube clamp 18 to cover the entire travel range from the centrifuge transfer position to the first centrifuge 5 adapter. It covers blood collection tubes with a height of 75-100mm; the double-speed belt mechanism can improve the motion control efficiency in the Z direction. After taking the blood collection tube from the material rack at the centrifuge transfer position, the blood collection tube is sent into the adapter installed on the first centrifuge 5. The total stroke is about 350mm and the time is less than 2s; the gripper actuator of the blood collection tube gripper 18 uses a parallel gripper actuator with a stroke of 10mm and a clamping force of 10mm. With the gripper, it can stably grip tubular objects up to 30g; the gripper adopts a 120° angle, which has been verified to have the characteristics of superior gripping stability; the gripper claw pattern adopts a horizontal V-shaped texture, which can effectively match the blood collection tube and increase friction.
[0047] Taking the material rack gripper drive mechanism 13 as an example, the free end stroke of the X-axis drive mechanism is 1000mm, the transmission mechanism is a ball screw, and the power motor is a 400W servo motor; the free end stroke of the Y-axis drive mechanism is 400mm, the transmission mechanism is a synchronous belt, and the power motor is a 57 closed-loop stepper motor; the free end stroke of the Z-axis drive mechanism is 550mm, the transmission mechanism is a ball screw module, and the power motor is a 100W servo motor with brake. It can cover the feeding drawer 4, the material rack plate 27 of the pipetting workstation, the centrifuge transfer position, and the flow cytometer 10 positions, which are compatible. The gripper actuator uses a rotary gripper actuator with a maximum clamping force of 100N, and the gripper actuator can rotate 360°. It is convenient to change the direction after clamping the 96-well plate, which helps to optimize the table layout and make the overall layout more compact.
[0048] In some examples, the two 3D moving mechanisms mentioned above share a single X-axis guide rail.
[0049] In some examples, a magnet is installed on the centrifuge transfer station. The magnet can attract and position the material rack placed on the centrifuge transfer station to prevent some of the blood collection tubes from being pulled up by the tag when the blood collection tube clamp 18 is pulling out the blood collection tube, thus avoiding equipment failure.
[0050] The blood collection tube clamp 18 in this embodiment of the invention is highly compatible. It can be used to transport 13mm and 16mm blood collection tubes, and its stroke can cover the height of blood collection tubes from 75mm to 100mm.
[0051] In some embodiments, the pipetting workstation includes a workstation gripper 19, a barcode scanner, a workstation gripper drive mechanism 20, a blood collection tube fixing clamp 21, a pipette 22, a pipette drive device 23, a perforated plate 24 placement position, and a recognition camera. A perforated plate 24 is placed at the perforated plate 24 placement position. The blood collection tube fixing clamp 21 can clamp and release the blood collection tube body. The recognition camera is located on one side of the blood collection tube fixing clamp 21 and is used to capture image information of the blood collection tube on the blood collection tube fixing clamp 21 and transmit it to the industrial control computer 3. The industrial control computer 3 determines the position information of the white blood cell layer inside the blood collection tube based on the image information. The workstation gripper 19 can clamp and release the blood collection tube body and cap on the pipetting workstation material rack plate 27. The workstation gripper drive mechanism... Mechanism 20 can drive the workstation gripper 19 to move, and the range of motion of the workstation gripper 19 covers the pipetting workstation material rack plate 27 and the blood collection tube fixing clamp 21. The workstation gripper driving mechanism 20 can also drive the workstation gripper 19 to rotate. The pipette driving device 23 can drive the pipette 22 to move, and the range of motion of the pipette 22 covers the placement position of the porous plate 24 and the blood collection tube fixing clamp 21. The industrial control computer 3 controls the pipette driving device 23 to move the inlet and outlet of the pipette 22 to the white blood cell layer according to the position information of the white blood cell layer in the blood collection tube to absorb the white blood cell layer. The barcode scanner is integrated with the workstation gripper 19 and is located on one side of the workstation gripper 19. The barcode scanner is used to scan the identification code on the blood collection tube.
[0052] This embodiment can scan the transported blood collection tubes, open the blood collection tube caps, identify the location of the white blood cell layer in each blood collection tube, extract the white blood cell layer and add it to the multi-well plate 24.
[0053] The specific process is as follows: After the material rack drive mechanism 30 drives the material rack plate 27 of the pipetting workstation to move into place in the pipetting workstation, the industrial control computer 3 controls the workstation gripper drive mechanism 20 to drive the workstation gripper 19 to move into place and clamp the cap of the blood collection tube, and pull the blood collection tube out of the material rack and transfer it to the blood collection tube fixing clamp 21 set on one side. During the transfer process, the material rack drive mechanism 30 can also drive the blood collection tube to rotate around its own axis. During the rotation, the barcode scanner continues to work so that the identification code on the blood collection tube can be successfully scanned when it is facing the barcode scanner. This scanning process does not require medical staff to place the blood collection tube in a specific orientation according to the rules, which improves the scanning success rate, the efficiency of manual operation and the fault tolerance. After the blood collection tube is placed on the blood collection tube fixing clamp 21, it is clamped and fixed in place. The workstation gripper drive mechanism 20 drives the workstation gripper 19 to rotate again and lift the tube cap, thus unscrewing the tube cap from the tube. After unscrewing, the tube cap is driven away from the tube to facilitate subsequent aspiration of the white blood cell layer. Before aspirating the white blood cell layer, the recognition camera captures a layered image inside the blood collection tube. The industrial control computer 3 controls the pipette drive device 23 to move the inlet and outlet of the pipette 22 to the white blood cell layer based on the position information of the white blood cell layer inside the blood collection tube to aspirate the white blood cells. After aspiration, the pipette drive device 23 drives the pipette 22 to move to the multi-well plate 24, and then the white blood cells in the pipette 22 are discharged into the wells of the multi-well plate 24. The industrial control computer 3 can also control the workstation gripper drive mechanism 20 to drive the workstation gripper 19 to carry the tube cap for capping, so that the complete blood collection tube can be returned to the feed drawer 4 later.
[0054] In some embodiments, multiple workstation grippers 19, barcode scanners, blood collection tube fixing clamps 21, and pipettes 22 are provided, preferably eight, so that eight blood collection tubes can be processed at a time. Multiple workstation grippers 19 are arranged side by side to form a multi-channel gripper; multiple barcode scanners are arranged side by side to form a multi-channel barcode scanner; multiple blood collection tube fixing clamps 21 are arranged side by side to form a multi-channel fixing clamp; multiple pipettes 22 are arranged side by side to form a multi-channel pipette 22; one workstation gripper 19 corresponds to one barcode scanner and one blood collection tube fixing clamp 21; one pipette 22 corresponds to one blood collection tube fixing clamp 21.
[0055] This embodiment achieves the goal of processing multiple blood collection tubes at once.
[0056] In some embodiments, the flow cytometry pretreatment mechanism includes a reagent storage mechanism, a temperature control module 26, an oscillation module 25, a second centrifuge 8, a multi-well plate gripper 28, and a multi-well plate gripper drive mechanism 29. The reagent storage mechanism is used to store the reagents required for pretreatment. The temperature control module 26 is used to perform temperature-controlled incubation of the sample in the multi-well plate 24. The oscillation module 25 is used to perform oscillation of the sample in the multi-well plate 24. The second centrifuge 8 is used to perform centrifugation of the sample in the multi-well plate 24. The stroke of the pipette 22 also covers the reagent storage mechanism. The multi-well plate gripper 28 can clamp and release the multi-well plate 24. The multi-well plate gripper drive mechanism 29 drives the multi-well plate gripper 28 to move and makes the stroke of the multi-well plate gripper 28 cover the placement position of the multi-well plate 24, the temperature control module 26, the oscillation module 25, and the second centrifuge 8.
[0057] This embodiment realizes the addition of reagents, centrifugation, shaking and temperature-controlled incubation of the multi-well plate 24.
[0058] Specifically, the pretreatment process in this embodiment includes: adding erythropoietin to multi-well plate 24 → vortexing for 10-20 seconds, incubating at 4°C for 10 minutes, centrifuging, discarding the supernatant, adding PBS to multi-well plate 24, vortexing for 10-20 seconds, centrifuging, discarding the supernatant, adding staining solution to multi-well plate 24 → vortexing for 10-20 seconds, incubating at 37°C for 30 minutes, centrifuging, discarding the supernatant, adding PBS to multi-well plate 24, vortexing for 10-20 seconds, centrifuging, discarding the supernatant, adding PBS to multi-well plate 24 and resuspending by pipetting, vortexing for 10-20 seconds, centrifuging, discarding the supernatant, adding PBS to multi-well plate 24 and resuspending by pipetting.
[0059] The processes of adding lecithin, PBS, staining solution, discarding supernatant, and resuspending by pipetting are implemented by pipette 22, the oscillation mixing is implemented by oscillation module 25, centrifugation is implemented by second centrifuge 8, and incubation is implemented by temperature control module 26.
[0060] In some embodiments, multiple pipettes 22 are arranged at equal intervals along the horizontal direction, each pipette 22 is vertically arranged, the inlet and outlet of the pipette 22 are constructed at the bottom of the pipette 22, and the pipette drive device 23 can drive each pipette 22 in the multi-channel pipette 22 to move up and down individually, and can change the position of all pipettes 22 in the multi-channel pipette 22 in the horizontal direction to achieve synchronous adjustment of the spacing between any two adjacent pipettes 22.
[0061] Because the hole spacing in the multi-well plate 24 and the axial distance between the blood collection tubes on the tray are different, the horizontal position of each pipette 22 in the multi-channel pipette 22 needs to be adjusted to adapt to the changes in hole spacing. For example, the center-to-center distance between the holes in the multi-well plate 24 is 9 mm, and the center-to-center distance between the blood collection tubes on the tray is 45 mm. The pipette 22 needs to adapt to the multi-well plate 24 with a center-to-center distance of 9 mm and the tray with a center-to-center distance of 45 mm. This requires that the spacing between all pipettes 22 be adjustable synchronously. Adjustable spacing multi-channel pipettes are a conventional method and will not be described in detail here.
[0062] Since the height of the white blood cell layer in each blood collection tube is not exactly the same, each pipette 22 is configured to be individually raised and lowered in order to aspirate white blood cells at different heights.
[0063] In some embodiments, the transfer mechanism 11 further includes a well plate support frame and a well plate support frame drive mechanism. The well plate support frame is used to carry the well plate 24. The stroke of the well plate gripper 28 covers the well plate support frame. The well plate support frame drive mechanism can drive the well plate support frame to move within the range of motion of the material rack gripper 17. The material rack gripper drive mechanism 13 can drive the material rack gripper 17 to clamp the well plate and send the well plate into the flow cytometer 10.
[0064] This embodiment achieves the purpose of feeding the pre-treated plate into the flow cytometer 10. The pipetting station rack plate 27 holds four racks at a time, and the 32 blood collection tubes on the four racks are arranged in four rows and eight columns, that is, eight blood collection tubes in each row. This facilitates the transfer of the white blood cell layer from the blood collection tubes on the pipetting station rack plate 27 to the multi-well plate 24.
[0065] In some embodiments, a UV disinfection module is provided inside the chassis 31, and part of the wall of the chassis 31 is constructed as a viewing window, which is made of UV-resistant organic glass; the chassis 31 is also provided with an H14 grade negative pressure HEPA filter module 12.
[0066] This embodiment prevents pathogen spillover.
[0067] In some embodiments, after the test is completed, the transfer mechanism 11 can return the blood collection tubes, the material rack, and the perforated plate 24 to their original positions.
[0068] This embodiment facilitates manual off-machine operation and avoids errors.
[0069] In some embodiments, the industrial control computer 3 automatically binds information to each blood collection tube, including the blood collection tube number and the test result information.
[0070] This embodiment requires no human intervention throughout the entire process, thus avoiding human error.
[0071] In some embodiments, the first centrifuge 5 and the second centrifuge 8 are disposed inside the casing 31 and positioned on the ground by a positioning frame 16 independent of the casing 31.
[0072] This embodiment can avoid the vibration of the first centrifuge 5 and the second centrifuge 8 affecting the accuracy of the white blood cell layer taken by the workstation.
[0073] In some embodiments, the chassis 31 is divided into a first outer shell 1 and a second outer shell 2. The opposite sides of the first outer shell 1 and the second outer shell 2 are both open structures. The open structures of the first outer shell 1 and the second outer shell 2 are joined together to form the chassis 31. A central support plate is fixedly provided in the middle of the first outer shell 1 and the second outer shell 2. The first centrifuge 5 and the flow cytometer 10 are disposed in the first outer shell 1 and located below the central support plate. The material rack gripper 17, the blood collection tube gripper 18, the material rack gripper drive mechanism 13, the blood collection tube gripper drive mechanism 14, the centrifuge transfer station, and the feed drawer 4 are all disposed in the first outer shell 1 and located above the central support plate. The pipetting workstation and the flow cytometry pretreatment mechanism are all disposed in the second outer shell 2 and located above the central support plate. The second centrifuge 8 is disposed in the second outer shell 2 and located below the central support plate. An operation window is provided on the central support plate. In the working state, the operation window is located directly above the second centrifuge 8 to facilitate the transfer of the perforated plate to the second centrifuge 8 through the operation window.
[0074] In this embodiment, the first outer shell 1 and the second outer shell 2 are detachably connected, which facilitates the disassembly and transportation of the entire system. Specifically, the first outer shell 1 and the second outer shell 2 are both 1800mm long, 850mm wide, and 1900mm high. After disassembly, they can enter passages, elevators, and doors with a width of 900mm or more.
[0075] In some examples, the first centrifuge 5 is an automated integrated centrifuge with the following performance characteristics: (1) automatic opening and closing of the chamber door; (2) automatic rotor positioning, which can be precisely stopped at a preset angle according to the program, facilitating automatic gripping by the equipment's blood collection tube grippers 18; and (3) software communication: it can automatically communicate with the equipment's host computer to achieve equipment linkage. The centrifugation parameters can achieve a speed of 0-5500 rpm, and the centrifugation temperature can reach a minimum of -20℃. The centrifuge rotor is a horizontal rotor with 4 adapters. Each adapter can hold 8 blood collection tubes, and a maximum of 32 blood collection tubes can be centrifuged at one time.
[0076] In some examples, the oscillation module 25 has an amplitude of ±1.5mm and a frequency of less than 1500rpm. The temperature control module 26 has a temperature control range of 0-99℃ and a temperature control accuracy of ±0.2℃.
[0077] In some examples, the multi-well plate 24 is a 96-well plate. The well arrangement is 12*8, meaning that the multi-channel gripper, multi-channel barcode scanner, multi-channel fixture, and multi-channel pipette 22 are all 8-channel.
[0078] In some examples, the second centrifuge 8 is an automated integrated centrifuge with the following performance characteristics: (1) automatic opening and closing of the chamber door; (2) automatic rotor positioning, which can be precisely stopped at a preset angle according to the program, facilitating automatic gripping of the 96-well plate by the equipment's grippers; (3) software communication: it can automatically communicate with the host computer of the equipment to realize equipment linkage. The centrifugation parameters can achieve a speed of 0-5500 rpm, and the centrifugation temperature can reach a minimum of -20℃. The centrifuge rotor is a horizontal rotor, containing 2 adapters. Each adapter can accommodate 1 96-well plate, and a maximum of 2 96-well plates can be centrifuged at a time.
[0079] In some examples, the flow cytometer 10 is branded as Sinopharm (Suzhou) Medical Technology Co., Ltd.; model number is BioCyte B4R2.
[0080] In some examples, the flow cytometer 10 can be fully pulled out via a two-stage drawer structure 15 for easy manual handling; the drawer structure includes an electromagnet hard limit to prevent the equipment from being pulled out during operation, which could lead to equipment failure and danger; the drawer includes a hydraulic buffer to cushion the impact and reduce noise when pushed in.
[0081] Detecting the expression of specific proteins using fluorescently labeled antibodies is the most common phenotypic identification method (e.g., using cytokeratin for positive identification, CD45 to exclude leukocytes, and DAPI staining of cell nuclei). Its advantage is that cells can be directly observed, but its disadvantages include time-consuming and labor-intensive image scanning and interpretation, and susceptibility to subjective factors. Currently, the enrichment and detection of CTCs based on fluorescent nanomaterials mainly relies on immunoaffinity; however, it has limitations in purity, capture efficiency, and sensitivity. In this embodiment of the invention, the flow cytometer 10 uses an active circulating tumor-associated rare cell staining solution to detect leukocytes. It recognizes tumor cells through cell membrane permeability, requires no antibody modification, and does not require immunoaffinity to recognize tumor cells, making the method simple and highly sensitive.
[0082] Specific examples have been used to illustrate the principles and implementation methods of this invention. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of this invention. Furthermore, those skilled in the art will recognize that, based on the ideas of this invention, there will be changes in the specific implementation methods and application scope. Therefore, the content of this specification should not be construed as a limitation of this invention.
Claims
1. A fully automated high-throughput blood testing system, characterized in that: include: Chassis; The feed drawer is equipped with a material rack for carrying blood collection tubes containing whole blood. The feed drawer is slidably mounted on the chassis to feed the blood collection tubes into or out of the chassis. The first centrifuge, located inside the casing, is used to centrifuge blood collection tubes containing whole blood. A transfer mechanism is installed inside the chassis. The transfer mechanism can transfer the blood collection tubes in the feed drawer to the first centrifuge; the transfer mechanism can also transfer the centrifuged blood collection tubes to the pipetting workstation. A pipetting workstation is installed inside the chassis. The pipetting workstation is equipped with a multi-well plate. The pipetting workstation is used to scan the transported blood collection tubes, open the blood collection tube caps, identify the position of the white blood cell layer in each blood collection tube, aspirate the white blood cell layer and add it to the multi-well plate, and close the blood collection tube caps. A flow cytometry pretreatment unit is located inside the chassis. The flow cytometry pretreatment unit is used to add reagents, centrifuge, shake, and incubate the multi-well plate containing a layer of white blood cells. A flow cytometer is installed inside the chassis. The transport mechanism is also used to transport the pretreated multi-well plate to the flow cytometer, which performs flow cytometry analysis and outputs the analysis results. An industrial control computer controls the operation of the first centrifuge, the transfer mechanism, the pipetting workstation, the flow cytometry pretreatment mechanism, and the flow cytometer.
2. The fully automated high-throughput blood testing system according to claim 1, characterized in that: The feeding drawer can accommodate 12 eight-hole material racks; the transfer mechanism includes material rack grippers, blood collection tube grippers, material rack gripper drive mechanism and blood collection tube gripper drive mechanism. The material rack gripper drive mechanism can drive the material rack grippers to transfer the material racks in the feeding drawer to the centrifuge transfer position. The blood collection tube gripper drive mechanism can drive the blood collection tube grippers to clamp the blood collection tubes in the material racks on the centrifuge transfer position into the first centrifuge and to clamp the centrifuged blood collection tubes out of the first centrifuge and put them back into the material racks on the centrifuge transfer position. The transfer mechanism also includes a pipetting workstation rack plate and a rack plate driving mechanism. The rack grippers can transport the rack carrying the centrifuged blood collection tubes to the pipetting workstation rack plate, and the rack plate driving mechanism drives the pipetting workstation rack plate carrying the rack to move into the pipetting workstation.
3. The fully automated high-throughput blood testing system according to claim 2, characterized in that: The pipetting workstation includes workstation grippers, a barcode scanner, a workstation gripper drive mechanism, a blood collection tube fixing clamp, a pipette, a pipette drive device, a multi-well plate placement position, and a recognition camera. A multi-well plate is placed at the multi-well plate placement position. The blood collection tube fixing clamp can clamp and release the blood collection tube body. The recognition camera is located on one side of the blood collection tube fixing clamp and is used to capture image information of the blood collection tube on the blood collection tube fixing clamp and transmit it to the industrial control computer. The industrial control computer determines the position information of the white blood cell layer within the blood collection tube based on the image information. The workstation grippers can clamp and release the blood collection tube body and cap on the material rack plate of the pipetting workstation. The workstation gripper drive mechanism can drive... The workstation gripper moves such that its range of motion covers the pipetting workstation material rack and the blood collection tube fixing clamp. The workstation gripper drive mechanism can also drive the workstation gripper to rotate. The pipette drive device can drive the pipette to move such that its range of motion covers the multi-well plate placement position and the blood collection tube fixing clamp. The industrial control computer controls the pipette drive device to move the inlet and outlet of the pipette to the white blood cell layer according to the position information of the white blood cell layer in the blood collection tube to aspirate the white blood cells. The barcode scanner is integrated with the workstation gripper and located on one side of the workstation gripper. The barcode scanner is used to scan the identification code on the blood collection tube.
4. The fully automated high-throughput blood testing system according to claim 3, characterized in that: The workstation grippers, barcode scanners, blood collection tube clamps, and pipettes are all provided in multiple quantities. Multiple workstation grippers are integrated side by side to form a multi-channel gripper; multiple barcode scanners are integrated side by side to form a multi-channel barcode scanner; multiple blood collection tube clamps are integrated side by side to form a multi-channel clamp; multiple pipettes are integrated side by side to form a multi-channel pipette; one workstation gripper corresponds to one barcode scanner and one blood collection tube clamp; one pipette corresponds to one blood collection tube clamp.
5. The fully automated high-throughput blood testing system according to claim 4, characterized in that: The flow cytometry pretreatment mechanism includes a reagent storage mechanism, a temperature control module, an oscillation module, a second centrifuge, multi-well plate grippers, and a multi-well plate gripper driving mechanism. The reagent storage mechanism is used to store the reagents required for pretreatment. The temperature control module is used to perform temperature-controlled incubation of the samples in the multi-well plate. The oscillation module is used to oscillate the samples in the multi-well plate. The second centrifuge is used to centrifuge the samples in the multi-well plate. The pipette stroke also covers the reagent storage mechanism. The multi-well plate grippers can clamp and release the multi-well plate. The multi-well plate gripper driving mechanism drives the multi-well plate grippers to move, so that the multi-well plate gripper stroke covers the multi-well plate placement position, the temperature control module, the oscillation module, and the second centrifuge.
6. The fully automated high-throughput blood testing system according to claim 4, characterized in that: Multiple pipettes are arranged at equal intervals along the horizontal direction, and each pipette is vertically arranged. The inlet and outlet of each pipette are located at the bottom of the pipette. The pipette driving device can individually drive each pipette in the multi-channel pipette to move up and down, and can change the horizontal position of all the pipettes in the multi-channel pipette to achieve synchronous adjustment of the spacing between any two adjacent pipettes.
7. The fully automated high-throughput blood testing system according to claim 5, characterized in that: The transfer mechanism further includes a well plate support frame and a well plate support frame drive mechanism. The well plate support frame is used to carry the well plate. The stroke of the well plate gripper covers the well plate support frame. The well plate support frame drive mechanism can drive the well plate support frame to move within the range of motion of the material rack gripper. The material rack gripper drive mechanism can drive the material rack gripper to clamp the well plate and send the well plate into the flow cytometer.
8. The fully automated high-throughput blood testing system according to claim 1, characterized in that: The chassis is equipped with an ultraviolet disinfection module, and part of the chassis wall is constructed as a viewing window, which is made of ultraviolet-resistant organic glass; the chassis is also equipped with an H14 grade negative pressure HEPA filter module.
9. The fully automated high-throughput blood testing system according to claim 5, characterized in that: The first centrifuge and the second centrifuge are housed inside the casing and positioned on the ground by a positioning frame independent of the casing.
10. The fully automated high-throughput blood testing system according to claim 5, characterized in that: The chassis is divided into a first outer shell and a second outer shell. The opposite sides of the first outer shell and the second outer shell are both open structures. The open structures of the first outer shell and the second outer shell are joined together to form the chassis. A central support plate is fixedly installed in the middle of both the first outer shell and the second outer shell. The first centrifuge and the flow cytometer are located inside the first outer shell and below the central support plate. The feed rack gripper, blood collection tube gripper, feed rack gripper drive mechanism, blood collection tube gripper drive mechanism, centrifuge transfer station, and feed drawer are all located inside the first outer shell and above the central support plate. The pipetting workstation and the flow cytometry pretreatment mechanism are all located inside the second outer shell and above the central support plate. The second centrifuge is located inside the second outer shell and below the central support plate.