A fully automatic flow cytometer
The design of a fully automated flow cytometer has achieved full automation of flow cytometer sample pretreatment and indiscriminate sample detection, solving the problems of high manual operation costs, large errors, and insufficient throughput, and improving the consistency of sample processing and detection efficiency.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- HANGZHOU CELLGENE BIOTECH CO LTD
- Filing Date
- 2025-07-21
- Publication Date
- 2026-06-02
AI Technical Summary
Existing flow cytometry detection equipment suffers from problems such as high manual operation, large errors, inconsistent sample processing, and insufficient throughput. In particular, it cannot achieve full automation in the sample pretreatment process and cannot simultaneously detect cells and proteins.
A fully automated flow cytometer was designed, comprising a sample loading module, a reagent tray module, a dilution tray module, a gripper arm assembly module, an incubation tray module, a magnetic separation module, a consumables tray module, a sample loading and disposal module, and a flow cytometry detection module. It automates the entire process of sample dilution, sample dispensing, reagent dispensing, isothermal incubation, reaction system cleaning, and flow cytometry detection, and is compatible with both cell and protein detection.
It achieves full automation of flow cytometry sample pretreatment, improves the consistency and reliability of sample processing, reduces detection variability, and enables simultaneous cell and protein detection, thereby improving detection efficiency.
Smart Images
Figure CN224317628U_ABST
Abstract
Description
Technical Field
[0001] This utility model belongs to the field of flow cytometry detection, and more specifically, relates to a fully automated flow cytometer. Background Technology
[0002] A flow cytometer, also known as a flow cell analyzer, is a multifunctional experimental instrument widely used in cell biology and molecular biology. It is primarily used for the quantitative analysis and sorting of cells. It integrates multiple technologies, including fluorescence, laser, monochromatic light, fluid dynamics, and computer technology. Through laser irradiation and fluorescence detection, it can rapidly and accurately analyze various cellular properties, including cell size, morphology, number, activity, and surface antigens. Furthermore, through specific fluorescent labeling methods, it can also perform quantitative and qualitative analysis of specific intracellular proteins.
[0003] Flow cytometry analysis has expanded to include disciplines such as cell biology, immunology, oncology, genetics, hematology, and microbiology. Current flow cytometry analysis faces several challenges: First, manual operation. Sample pretreatment still requires manual intervention, resulting in high labor costs and a heavy workload for operators. Second, controlling operational errors is difficult. Error rates increase with operation time, such as missed or incorrect sample additions, compromising sample consistency. Furthermore, manual sample processing poses biosafety risks to operators. Finally, insufficient throughput. Purely manual sample preparation limits the ability to prepare large batches of samples, resulting in low sample processing rates and hindering the development of flow cytometry platforms.
[0004] Therefore, with the increasing demand for flow cytometry, more and more flow cytometry devices capable of sample pretreatment have emerged on the market. For example, the device disclosed in patent CN107422138A does not completely eliminate all manual operations. It still requires manual mixing of the sample to be tested (blood sample in a vacuum blood collection tube) and placement of the blood sample onto the blood sample loading position using a pipette. Operators, especially those in disease control centers, still have to handle high-risk, easily infectious samples, exposing them to potential biological risks. Furthermore, this device does not automate the mixing and transfer of the sample to be tested; it is only a semi-automated instrument operation. Another example is the multi-mode flow cytometry lysis-elution pretreatment system disclosed in patent CN222979384U. While it can perform cell flow cytometry pretreatment and microsphere flow cytometry pretreatment, it lacks a flow cytometer and cannot directly analyze the pretreated sample. Currently, there are almost no fully automated flow cytometers on the market that can perform both protein and cell detection. Most fully automated flow cytometers with fluorescence immunoassay can only perform protein detection and cannot perform cell detection; cell detection still relies on traditional flow cytometers. Some manufacturers use a combination of different instruments, but these are not truly fully automated.
[0005] Therefore, there is an urgent need for a fully automated flow cytometer that not only frees up manual labor but also minimizes errors and maximizes consistency in sample processing. The entire process from sample collection to processing is automated and intelligently controlled, truly achieving "sample in, result out" and improving the throughput of flow cytometers (TAT) in the laboratory. Utility Model Content
[0006] To address the challenges of existing technologies, this invention provides a fully automated flow cytometer, comprising a sample loading module, a reagent tray module, a reagent needle module, a sample loading module, a dilution tray module, a gripper arm assembly module, an incubation tray module, a magnetic separation module, a consumables tray module, a sample loading and disposal module, and a flow cytometry detection module. These modules work together to complete the entire flow cytometry process, including sample dilution, sample dispensing, reagent dispensing, isothermal incubation, reaction system cleaning, and flow cytometry detection. This fully automated flow cytometer can automatically complete sample pretreatment and detection, improving the consistency and reliability of sample pretreatment and reducing detection errors. It is compatible with both protein biomarkers and cell detection, supporting seamless operation for both protein and cell analysis. The instrument automatically identifies the detection item, grabs the corresponding detection tube, and executes the appropriate program, eliminating the need for manual differentiation and improving detection efficiency.
[0007] This invention provides a fully automated flow cytometer, including a dilution tray module. The dilution tray module includes a dilution tray and six working positions. The dilution tray is provided with multiple insertion holes that can rotate in place, and the insertion holes are used to place reaction tubes. The six working positions include a first reaction tube pick-up / placement position, a diluent addition position, a sample addition position, a reagent addition position, a vortexing and mixing position, and a waste liquid removal and tube discarding position. During the rotation of the dilution tray, the insertion holes can be rotated to the six working positions.
[0008] The sample enters the fully automated flow cytometer through the sample entry module, and then enters the reaction tube of the dilution tray through the sample loading module for sample pretreatment, including sample dilution and sample-reagent mixing. Since the dilution tray is rotatable, and 6 working positions are set below the dilution tray, each socket can be rotated to 6 different positions for different sample treatments during the dilution tray rotation. The specific process of sample dilution and reagent addition in the dilution tray is as follows: (1) Sample loading: The grab arm module grabs an empty reaction tube and places it into the first reaction tube pick / place position. Then the dilution tray rotates to the sample loading position, and the sample loading module picks up the sample and puts it into the reaction tube; (2) Dilution: The dilution tray rotates to the diluent loading position, and the reagent needle module picks up the diluent and adds it into the reaction tube. The dilution tray rotates to the oscillation and mixing position to mix. The sample loading module picks up the diluted sample in the reaction tube and keeps it in the picking state. The dilution tray rotates to the waste liquid removal and tube discarding position to grab the waste reaction tube after the diluted sample is picked up, and to remove the waste liquid. Discard the tube, rotate the dilution tray to the first reaction tube pick / place position, the gripper arm module picks up a new empty reaction tube and places it into the same socket, the sample addition module adds the aspirated diluted sample to the new empty reaction tube. If multiple dilutions are required, repeat the dilution process; (3) Add reagent: the dilution tray rotates the reaction tube to the reagent addition position, the reagent needle module picks up the reagent from the reagent tray module and adds it to the reaction tube, the dilution tray rotates to the oscillation mixing position for mixing; (4) Transfer the reaction tube: the dilution tray rotates to the first reaction tube pick / place position and waits for the gripper arm module to pick up the reaction tube for the next operation. If the sample does not need to be diluted, the dilution tray does not perform the dilution step during rotation and directly rotates to the reagent addition position.
[0009] Furthermore, the fully automated flow cytometer also includes a sample entry module, a sample addition module, a reagent needle module, and a reagent tray module; the reagent needle module includes a reagent needle, a reagent needle moving mechanism, and a needle washing pool, and the reagent needle moving mechanism drives the reagent needle to move between the reagent tray module, the dilution tray module, and the needle washing pool.
[0010] The process of adding diluent or reagent to the reaction tube in the dilution pan socket is completed through the reagent needle module.
[0011] In some embodiments, the reagent needle moving mechanism drives the reagent needle to move in an arc around a rotation axis. The reagent tray module, dilution tray module, and needle washing pool surround the reagent needle module, so the reagent needle moving mechanism drives the reagent needle to move between the reagent tray module, dilution tray module, and needle washing pool.
[0012] Furthermore, the sample entry module includes a sample holder, a push-in area, and a transverse axis, wherein the push-in area and the transverse axis can drive the sample holder to move.
[0013] Place the sample tube (such as a blood collection tube or EP tube) into the sample rack. Tracks are set on both sides of the push-in area. The sample rack is placed horizontally into the push-in area. The track in the push-in area sends the sample rack into the horizontal movement axis. The horizontal movement axis can drive the sample rack to move horizontally into the junction of the sample entry module and the sample dispensing module.
[0014] Furthermore, the sample entry module also includes a sample tube identification device, a sample identification device, a shaking device, and a sample tube unloading device.
[0015] The sample tube identification device is equipped with a sensor to identify the presence / absence of a sample tube and the presence / absence of a tube cap; the sample identification device can rotate the sample tube and scan the code for identification; the shaking device clamps the sample tube and shakes it; the sample unloading device pushes the sample tube out of the sample rack and into the sample entry module.
[0016] Furthermore, the sample loading module includes a loading needle, a loading needle moving mechanism, and a loading needle cleaning mechanism. The loading needle and the loading needle moving mechanism are located above the dilution tray module and the sample entry module. The loading needle moving mechanism drives the loading needle to move between the dilution tray module and the sample entry module.
[0017] The sample loading module has two main functions: sampling and loading, and transferring the diluted sample from the reaction tube already in the dilution pan socket to a new empty reaction tube located adjacent to the socket in the dilution pan. During sampling, the loading needle moving mechanism moves the loading needle above the sample entry module to puncture and sample the sample from the tube. During loading, the loading needle moving mechanism moves the loading needle to the loading position in the dilution pan to add the sample.
[0018] The sample needle cleaning mechanism cleans the needle by lifting the sample needle.
[0019] Furthermore, the reagent tray module includes a refrigerated reagent position and a room temperature reagent position. The refrigerated reagent position includes a reagent tray and a diluent bottle placement position, and the room temperature reagent position includes a hemolysin placement position. The reagent tray is equipped with a turntable, which has a conventional reagent placement position and a microsphere reagent placement position. The conventional reagent placement position is located on the outer ring of the turntable, and the microsphere reagent placement position is located on the inner ring of the turntable. The turntable performs reciprocating rotational motion. A mixing component is also provided at the bottom of the microsphere reagent placement position.
[0020] The microsphere reagents include magnetic microsphere reagents and non-magnetic microsphere reagents.
[0021] In the reagent tray module, the refrigerated reagent position is the refrigerated area. The reagent tray and diluent bottle placement position are located in the refrigerated area. The reagent tray is used to store conventional reagents and microsphere reagents, and the diluent bottle placement position is used to place diluent bottles and store diluents. The reagents stored here are stored under refrigerated conditions. The room temperature reagent position is the room temperature area. The hemolysin placement position is located in the room temperature area. It is used to place hemolysin bottles and store hemolysin. The hemolysin stored here is stored under room temperature conditions.
[0022] In the reagent tray, the turntable reciprocates around its axis to meet the requirements of the reagent needle to draw up different reagents; at the same time, a mixing component is set at the bottom of the microsphere reagent placement position on the turntable to mix the microsphere reagents stored there.
[0023] In some embodiments, the mixing component at the bottom of the microsphere reagent placement position is a small turntable that reciprocates, thus allowing the microsphere reagent stored thereto to be mixed.
[0024] In some embodiments, in the reagent tray module, the outer ring of the reagent tray has 16 conventional reagent placement positions, the inner ring of the reagent tray has 4 microsphere reagent placement positions, the number of diluent bottle placement positions is 1, and the number of hemolysin bottle placement positions is 1.
[0025] In some embodiments, the microsphere reagent is a magnetic microsphere reagent.
[0026] Furthermore, the fully automated flow cytometer also includes an incubation tray module, a magnetic separation module, a consumable tray module, a gripper arm assembly module, and a sample loading and discarding module; the incubation tray module includes an incubation tray with multiple incubation holes.
[0027] The incubation dish provides a suitable reaction temperature for the sample in the incubation reaction tube, and the entire dish oscillates in an eccentric manner to prevent microspheres from settling.
[0028] In some methods, the incubation tray has 150 incubation holes.
[0029] Furthermore, the magnetic separation module includes a magnetic separation disk that can rotate in place. The magnetic separation disk is provided with multiple magnetic separation holes, and a reaction tube is placed in each magnetic separation hole. The magnetic separation disk is also provided with 6 working positions, including a second reaction tube pick-up / placement position, a 1mL PBS addition position, a mixing position, a magnetic adsorption position, a waste liquid extraction position, and a 100mL PBS addition position. During the rotation of the magnetic separation disk in place, the magnetic separation holes can rotate to the 6 working positions.
[0030] The magnetic separation module is used to clean the samples after the reaction. Each reaction tube rotates once in the magnetic separation disk to complete one cleaning action. After incubation in the incubation tray, the samples in the reaction tubes are transferred to the magnetic separation disk by the grab arm module to complete one cleaning process as follows: The reaction tube is placed in the second reaction tube pick / place position; the magnetic separation disk rotates to the 1mL PBS addition position to add 1mL PBS; the magnetic separation disk rotates to the mixing position to mix; the magnetic separation disk rotates to the magnetic adsorption position and the waste liquid extraction position (the magnetic adsorption position and the waste liquid extraction position are located in the same position) for magnetic adsorption and supernatant extraction; the magnetic separation disk rotates to the 100mL PBS addition position to add 100mL PBS. If n cleanings are required, the magnetic separation disk repeats the operation of rotating to the 1mL PBS addition position, mixing position, magnetic adsorption position, and waste liquid extraction position n times, and after the last supernatant extraction, the magnetic separation disk rotates to the 100mL PBS addition position to add 100mL PBS.
[0031] In some methods, the magnetic separation disk rotates counterclockwise twice to complete two (second-stage) cleaning cycles.
[0032] The fully automated flow cytometer provided by this invention can detect both proteins (such as cytokines) and cells (such as lymphocyte subsets) in a sample. When the fully automated flow cytometer detects proteins (such as cytokines) in a sample, the aforementioned magnetic separation module is required. However, when detecting cells (such as lymphocyte subsets) in a sample, the magnetic separation module is not required. Therefore, the gripper arm module will grab the reaction tube, bypassing the magnetic separation module, and directly enter the sample loading and discarding module.
[0033] In some configurations, the magnetic separation module includes 13 stations: one each for taking / placing a second reaction tube, adding 1 mL of PBS, mixing, removing waste liquid, and adding 100 mL of PBS, and eight magnetic adsorption stations.
[0034] Furthermore, the consumables tray module is provided with multiple reaction tube storage positions; the gripper arm assembly module has a gripper and a gripper moving mechanism, and after the reaction tube is gripped by the gripper, it is transferred between the consumables tray module, dilution tray module, incubation tray module, magnetic separation module and sample loading and discarding module through the gripper moving mechanism.
[0035] The reaction tubes (such as flow cytometers, absolute counters, etc.) are stored on the reaction tube rack.
[0036] In some configurations, four reaction tube racks are set up, each rack can hold 30 reaction tubes, for a total of 120 reaction tubes.
[0037] Furthermore, the consumables tray module can also perform CCD recognition to photograph and identify the status of the reaction tubes in the module (ordinary empty tube / absolute counting tube / no tube / with or without tube cap).
[0038] Furthermore, the sample loading and discarding module includes a sample loading and discarding tray, which is rotatable in place. The sample loading and discarding tray is provided with multiple sample loading and discarding holes, in which reaction tubes are placed. The sample loading and discarding tray is also provided with four working positions, including a third reaction tube pick-up / placement position, a discard pick-up / placement position, a sample loading liquid pick-up position, and a waste liquid extraction position. During the rotation of the sample loading and discarding tray in place, the sample loading and discarding holes can rotate to the four working positions.
[0039] After the sample in the reaction tube completes the pretreatment process, it is grasped by the grabbing arm module and placed into the sample loading and disposal tray. First, the reaction tube enters the third reaction tube pick-up / placement position, and the sample loading and disposal tray rotates 180° to the sample loading position. The sampling needle draws the sample from the reaction tube and sends it to the flow cytometry detection module for analysis. If no retest is required, the sample loading and disposal tray rotates to the waste liquid removal position, and the waste liquid needle removes the waste liquid from the reaction tube. Then, the sample loading and disposal tray rotates back to the disposal pick-up / placement position, and the waste reaction tube automatically falls into the waste bin. If a retest is required, the sample loading and disposal tray rotates 180° back to the third reaction tube pick-up / placement position. The gripper arm module transfers the reaction tube back to the incubation tray. The gripper arm module then transfers the reaction tube from the incubation tray back to the third reaction tube pick-up / placement position on the sample loading and disposal tray. The sample loading and disposal tray rotates 180° to rotate the reaction tube back to the sample loading and liquid collection position. The aspiration needle draws up the sample and enters the flow cytometry detection module for re-inspection. Then, the sample loading and disposal tray rotates to the waste liquid collection position, and the waste liquid needle draws up the waste liquid in the reaction tube. Subsequently, the sample loading and disposal tray rotates back to the disposal pick-up / placement position, and the waste reaction tube automatically falls into the waste bin.
[0040] Furthermore, the fully automated flow cytometer also includes a flow cytometry detection module and a rack module.
[0041] The results detected by the streaming detection module will be displayed synchronously on the user interface.
[0042] The rack module provides support for the fully automated flow cytometer and space for the placement of each module.
[0043] This utility model has the following beneficial effects:
[0044] 1. This utility model can automatically complete the pretreatment and detection of flow cytometry samples. It consists of a sample entry module, a reagent tray module, a reagent needle module, a sample addition module, a dilution tray module, a gripper arm assembly module, an incubation tray module, a magnetic separation module, a consumable tray module, a sample loading and disposal module, a flow cytometry detection module, and a rack module. The modules work together to realize the entire process of flow cytometry experiments, including sample dilution, diluted sample distribution, reagent distribution, magnetic adsorption, mixing, and incubation.
[0045] 2. This utility model is compatible with protein and cell detection. It is a truly fully automated flow cytometer that allows for "sample in, result out". After the sample enters the instrument, it undergoes automated pretreatment and automated detection.
[0046] 3. This invention can be used without discrimination and can process cell and protein detection reagents simultaneously. Cell detection (such as lymphocyte subsets) and protein detection items can be processed at the same time. The instrument can automatically identify the detection items, grab the corresponding detection tubes, and execute different programs without the need for manual differentiation. Attached Figure Description
[0047] Figure 1 Schematic diagram of the overall structure of a fully automated flow cytometer Figure 1 (From left to right, from front to back);
[0048] Figure 2 Schematic diagram of the overall structure of a fully automated flow cytometer Figure 2 (From right to left, from back to front);
[0049] Figure 3 A top view of a fully automated flow cytometer;
[0050] Figure 4 This is a top view of the reagent tray module;
[0051] Figure 5 Left view of an automated flow cytometer;
[0052] Figure 6 This is a front view of an automated flow cytometer.
[0053] Figure 7 This is a right front oblique view of an automated flow cytometer.
[0054] Figure 8 Right view of an automated flow cytometer;
[0055] Figure 9 Rear view of an automated flow cytometer;
[0056] Figure 10 This is a left rear oblique view of an automated flow cytometer.
[0057] Figure 11 This is a right rear oblique view of an automated flow cytometer. Detailed Implementation
[0058] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be noted that the embodiments described below are intended to facilitate the understanding of the present invention and do not constitute any limitation thereof.
[0059] Example 1: A fully automated flow cytometer
[0060] like Figure 1 and 2 As shown, this utility model provides a fully automated flow cytometer, comprising a sample entry module 1, a reagent tray module 2, a reagent needle module 3, a sample loading module 4, a dilution tray module 5, a gripper arm assembly module 6, an incubation tray module 7, a magnetic separation module 8, a consumable tray module 9, a sample loading and discarding module 10, a flow cytometry detection module 11, and a frame module 12.
[0061] like Figure 1-11 As shown, the sample entry module 1 includes a sample rack 13, a push-in area 14, and a transverse axis 15. The sample rack 13 is used to hold sample tubes, including but not limited to blood collection tubes and EP tubes. Tracks are provided on both sides of the push-in area 14. The sample rack 13 is placed horizontally into the push-in area 14, and the push-in area 14 sends the sample rack 13 into the transverse axis 15. The transverse axis 15 can drive the sample rack 13 to move horizontally into the junction of the sample entry module 1 and the sample loading module 4. The sample entry module 1 also includes a sample tube identification device, a sample identification device, a shaking device, and a sample tube unloading device. The sample tube identification device is equipped with a sensor to identify the presence / absence of a sample tube and the presence / absence of a tube cap. The sample identification device can rotate the sample tube and scan the code for identification. The shaking device clamps the sample tube and shakes it. The sample unloading device pushes the sample tube out of the sample rack from the sample entry module 1.
[0062] The reagent tray module 2 includes a refrigerated reagent compartment and a room temperature reagent compartment. The refrigerated reagent compartment includes a reagent tray 16 and a diluent bottle placement compartment 49, while the room temperature reagent compartment includes a hemolysin placement compartment 17. The refrigerated reagent compartment, also known as the refrigerated zone, houses the reagent tray 16 and the diluent bottle placement compartment 49. The reagent tray 16 stores conventional reagents and microsphere reagents (including magnetic and non-magnetic microsphere reagents). There is one diluent bottle placement compartment 49, used to place diluent bottles 55 and store diluent. Reagents stored here are refrigerated. The room temperature reagent compartment, also known as the room temperature zone, houses one hemolysin placement compartment 17, used to place hemolysin bottles 54 and store hemolysin. Hemolysin stored here is at room temperature. The reagent tray 16 includes a turntable 50. The outer ring of the turntable 50 has 16 conventional reagent placement compartments 52, and the inner ring of the turntable 50 has 4 microsphere reagent placement compartments 53. The turntable 50 reciprocates around its axis to meet the requirements of the reagent needle for drawing different reagents. A mixing component 51 is installed at the bottom of the four microsphere reagent placement positions 53 to mix the microsphere reagents. The mixing component 51 is a small turntable that reciprocates, thus ensuring that the microsphere reagents stored there are mixed evenly.
[0063] The reagent needle module 3 is used to draw reagents (conventional reagents, microsphere reagents, and hemolysin) from the reagent tray module 2 and add them to the dilution tray module 5. The reagent needle module 3 includes a reagent needle 18, a reagent needle moving mechanism 19, and a needle washing basin. The reagent needle moving mechanism 19 drives the reagent needle 18 to move in an arc around a rotating axis 20. Since the reagent tray module 2, dilution tray module 5, and needle washing basin surround the reagent needle module 3, the reagent needle moving mechanism 19 moves the reagent needle 18 between these components.
[0064] The dilution tray module 5 includes a dilution tray 21, which can rotate in place and has multiple sockets for placing reaction tubes, which serve as containers for sample pretreatment. The dilution tray 21 has six different positions below it, and the sockets can rotate to six different positions during rotation, including a first reaction tube pick / place position 22, a diluent addition position 23, a sample addition position 24, a reagent addition position 25, a vortexing and mixing position 26, and a waste liquid removal and tube discarding position 27. The specific process of diluting the sample and adding reagents in the dilution tray 21 is as follows: (1) Sample addition: The grab arm module 6 grabs an empty reaction tube and puts it into the first reaction tube pick / place position 22. Then the dilution tray 21 rotates to the sample addition position 24, and the sample addition module 4 picks up the sample and puts it into the reaction tube; (2) Dilution: The dilution tray 21 rotates to the diluent addition position 23, and the reagent needle module 3 picks up the diluent and adds it into the reaction tube. The dilution tray 21 rotates to the oscillation and mixing position 26 to mix. The sample addition module 4 picks up the diluted sample in the reaction tube and keeps it in the aspiration state. The dilution tray 21 rotates to the waste liquid removal and tube discarding position 27 to grab the waste reaction tube after the diluted sample has been taken and remove the waste liquid. (2) Discarding the tube: The dilution tray 21 rotates to the first reaction tube pick / place position 22, the gripper arm module 6 picks up a new empty reaction tube and places it into the same socket 22, the sample addition module 4 adds the aspirated diluted sample to the new empty reaction tube. If multiple dilutions are required, the dilution process is repeated; (3) Adding reagents: The dilution tray 21 rotates the reaction tube to the reagent addition position 25, the reagent needle module 3 aspirates the reagent from the reagent tray module 2 and adds it to the reaction tube, the dilution tray 21 rotates to the oscillation mixing position 26 for mixing; (4) Transferring the reaction tube: The dilution tray 21 rotates to the first reaction tube pick / place position 22 and waits for the gripper arm module 6 to pick up the reaction tube for the next operation. If the sample does not need to be diluted, the dilution tray 21 does not perform the dilution step during rotation and directly rotates to the reagent addition position 25. The reagent tray module 5 is also equipped with a reagent tray air duct 44.
[0065] The sample loading module 4 includes a loading needle 28, a loading needle moving mechanism 29, and a loading needle cleaning mechanism. The loading needle 28 and the loading needle moving mechanism 29 are located above the dilution tray module 5 and the sample entry module 1. The loading needle moving mechanism 29 moves the loading needle 28 between the dilution tray module 5 and the sample entry module 1. The sample loading module 4 has two main functions: one is to aspirate the sample from the sample tube and add it to the reaction tube in the dilution tray 21; the other is to aspirate the diluted sample from the reaction tube in the dilution tray 21 and transfer it to a new empty reaction tube. When aspirating the sample from the sample tube, the loading needle moving mechanism 29 moves the loading needle 28 above the sample tube in the sample entry module 1 to puncture and sample the sample from the sample tube. When adding the aspirated sample to the reaction tube in the dilution tray 21, the loading needle moving mechanism 29 moves the loading needle 28 to the loading position 24 in the dilution tray 21 to add the sample. The loading needle cleaning mechanism cleans the needle by lifting the loading needle 28.
[0066] The incubation tray module 7 includes an incubation tray 30, which has multiple incubation holes. In this embodiment, the number of incubation holes is 150. The incubation tray 30 provides a suitable reaction temperature for the sample in the reaction tube. The entire tray oscillates in an eccentric manner to prevent microsphere sedimentation. An incubation tray water tank air duct 45 is provided below the incubation tray 30.
[0067] The magnetic separation module 8 includes a magnetic separation disk 31 and six working positions. The magnetic separation disk 31 is rotatable and has multiple magnetic separation holes, each containing a reaction tube. The six working positions include a second reaction tube pick-up / placement position, a 1mL PBS addition position, a mixing position, a magnetic adsorption position, a waste liquid removal position, and a 100mL PBS addition position. During the rotation of the magnetic separation disk 31, the magnetic separation holes can rotate to these six positions to complete various actions. The magnetic separation module 8 is used for sample cleaning; each reaction tube completes one cleaning cycle by rotating once within the magnetic separation disk 31. After incubation in incubation tray 30, the sample in the reaction tube is transferred by the grab arm module 6 to the magnetic separation tray 31 to complete one cleaning cycle as follows: The reaction tube is placed in the second reaction tube pick / place position; the magnetic separation tray 31 rotates to the 1mL PBS addition position to add 1mL PBS; the magnetic separation tray rotates to the mixing position to mix; the magnetic separation tray 31 rotates to the magnetic adsorption position and the waste liquid extraction position (the magnetic adsorption position and the waste liquid extraction position are located in the same position) for magnetic adsorption and supernatant aspiration; the magnetic separation tray 31 rotates to the 100mL PBS addition position to add 100mL PBS. If n cleaning cycles are required, the magnetic separation tray 31 repeats the rotation to the 1mL PBS addition position, mixing position, magnetic adsorption position, and waste liquid extraction position n times, completing each action, and after the last supernatant aspiration, the magnetic separation tray 31 rotates to the 100mL PBS addition position to add 100mL PBS. In this embodiment, there are one each for the second reaction tube pick-up / placement position, the 1mL PBS addition position, the mixing position, the waste liquid removal position, and the 100mL PBS addition position, and eight magnetic adsorption positions, for a total of 13 working positions. The fully automated flow cytometer provided by this utility model can detect both proteins (such as cytokines) and cells (such as lymphocyte subsets) in a sample. When the fully automated flow cytometer detects proteins (such as cytokines) in a sample, the aforementioned magnetic separation module 8 is required. However, when it detects cells (such as lymphocyte subsets) in a sample, the aforementioned magnetic separation module 8 is not required. Therefore, the gripper arm module 6 will grab the reaction tube, skip the magnetic separation module 8, and directly enter the sample loading and discarding module 10.
[0068] The consumables tray module 9 is used to store empty reaction tubes (such as flow cytometry tubes, absolute counter tubes, etc.), and is provided with multiple reaction tube storage positions 32, which are set on reaction tube racks 33. In this embodiment, four reaction tube racks are provided, each rack holding 30 reaction tubes, for a total of 120 reaction tubes. The consumables tray module 9 can also perform CCD recognition to electronically identify the status of the reaction tubes in the module (ordinary empty tubes / absolute counter tubes / no tubes / with or without tube caps).
[0069] The sample disposal module 10 includes a sample disposal tray 34 and four working positions. The sample disposal tray 34 can rotate in place and has multiple sample disposal holes inside, in which reaction tubes are placed. The four working positions include the third reaction tube pick / place position, the disposal pick / place position, the sample liquid collection position, and the waste liquid collection position. During the rotation of the sample disposal tray 34, the sample disposal holes can rotate to the four working positions. After the sample in the reaction tube completes the pretreatment process, it is picked up by the gripping arm module 6 and placed into the sample disposal tray 34. First, the reaction tube enters the third reaction tube pick / place position, the sample disposal tray rotates 180° to the sample liquid collection position, the aspiration needle draws the sample from the reaction tube into the lost cell detection module 11 for detection, the sample disposal tray 34 rotates to the waste liquid collection position, the waste liquid needle draws away the waste liquid in the reaction tube, and then the sample disposal tray 34 rotates back to the disposal pick / place position, and the waste reaction tube automatically falls into the waste bin 33.
[0070] The gripper assembly module 6 includes a gripper and a gripper moving mechanism. After the reaction tube is gripped by the gripper, it is transferred between the consumable tray module 9, the dilution tray module 5, the incubation tray module 7, the magnetic separation module 8, and the sample loading and discarding module 10 via the gripper moving mechanism.
[0071] The flow cytometry detection module 11 is used to detect samples, and the detection results are displayed synchronously on the user interface. The flow cytometry detection module 11 includes a detection module chassis, a detection module liquid channel area 42, and a detection module optical platform 43.
[0072] The rack module 12 provides support for the fully automated flow cytometer and space for the placement of various modules. Below the rack module 12 are multiple reagent tanks for storing reagents used in the detection process, waste reagents, and reagents used for maintenance and cleaning, including a PBS tank 36, a sheath fluid tank 37, a waste tank 33, a waste liquid tank 39, a cleaning solution tank 38, an alcohol bottle 40, and a maintenance solution bottle 41. Below the rack module 12 are also a liquid path area 46, a circuit board chassis 35, and an electrical area 48. The circuit board chassis 35 has a circuit board chassis exhaust vent 47.
[0073] The fully automated flow cytometer provided by this invention is a highly automated cell / protein analysis device that integrates sample processing, detection, and data analysis into one unit. It can simultaneously detect soluble proteins and cells, greatly simplifying experimental operations and improving analytical efficiency. Regarding sample processing, the fully automated flow cytometer can automatically complete the entire process, including sample dilution, diluted sample distribution, reagent dispensing, magnetic separation and washing, mixing, and incubation, without manual intervention. This significantly reduces the possibility of human error during experiments and improves the consistency and reliability of sample processing.
[0074] Example 2, Protein Detection Procedure
[0075] like Figure 3 As shown, taking the detection of cytokines as an example, the detection process using the fully automated flow cytometry analyzer provided by this invention is as follows:
[0076] 1. First, turn on the instrument. After powering on, the instrument will perform an initialization self-test. After initialization, the instrument is ready. Place the reagents needed for the experiment into the reagent slots of the reagent tray (place magnetic microsphere reagents in microsphere reagent slot 53), and place the reaction tubes (such as flow cytometers) into the consumables tray module 9. Other liquids have been prepared in advance (before initialization). Then, you can select and set the corresponding execution program in the software interface.
[0077] 2. Place the sample tube into the sample rack 13. The sample entry module 1 sends the sample rack 13 to the horizontal axis 15 through the push-in area 14. During this process, the sample entry module 1 identifies the sample tube and performs rotation scanning.
[0078] 3. The gripper of the gripper arm module 6 takes an empty reaction tube from the consumable tray module 9 and places it into the first reaction tube pick / place position 22 of the dilution tray 21. The dilution tray 21 is rotated to move the reaction tube to the sample addition position 24.
[0079] 4. After the sample needle 28 takes a sample from the sample tube in the test tube rack 13 pushed horizontally by the horizontal movement axis 15, it adds the sample to the sample addition position 24 of the dilution tray 21.
[0080] 5. Rotate the dilution tray 21 to move the reaction tube to the reagent addition position 25;
[0081] 6. The reagent needle 18 takes magnetic microsphere reagent from the microsphere reagent placement position 53 in the inner ring of the turntable 50 and microsphere buffer solution from the conventional reagent placement position 52 in the outer ring of the turntable 50 and adds it to the reagent addition position 25 in the dilution plate 21.
[0082] 7. Rotate the dilution tray 21 to move the reaction tube to the oscillation mixing position 26, and oscillate the sample and reagent to mix.
[0083] 8. Rotate the dilution tray 21 to move the reaction tube to the first reaction tube pick / place position 22;
[0084] 9. The gripper of the gripper arm module 6 transfers the reaction tube to the incubation tray 30 for incubation;
[0085] 10. The gripper of the gripper arm module 6 transfers the reaction tube back from the incubation plate 30 to the first reaction tube pick-up / placement position 22 of the dilution plate 21. The dilution plate 21 rotates until the reagent addition position 25. The reagent needle 18 takes the fluorescent detection reagent from the conventional reagent placement position 52 on the outer ring of the turntable 50 and adds it to the reaction tube located at the reagent addition position 25. Then the dilution plate 21 rotates to the oscillation mixing position 26 to mix and then rotates to the first reaction tube pick-up / placement position 22.
[0086] 11. The gripper of the gripper arm module 6 transfers the reaction tube back from the dilution tray 21 to the incubation tray 30 for shaking incubation;
[0087] 12. After incubation, the gripper of the gripper arm module 6 will transfer the reaction tube to the second reaction tube pick-up / placement position of the magnetic separation disk 31;
[0088] 13. Rotate the magnetic separation disk 31 counterclockwise 2 times to complete the second-stage cleaning: Rotate the magnetic separation disk 31 to the 1mL PBS addition position to add 1mL PBS, rotate the magnetic separation disk to the mixing position to mix, rotate the magnetic separation disk 31 to the magnetic adsorption position and waste liquid extraction position to perform magnetic adsorption and supernatant extraction, rotate the magnetic separation disk 31 to the 1mL PBS addition position to add 1mL PBS, rotate the magnetic separation disk to the mixing position to mix, rotate the magnetic separation disk 31 to the magnetic adsorption position and waste liquid extraction position to perform magnetic adsorption and supernatant extraction, rotate the magnetic separation disk 31 to the 100mL PBS addition position to add 100mL PBS, rotate the magnetic separation disk 31 to the second reaction tube dispensing / placing position;
[0089] 14. The gripper of the gripper arm module 6 transfers the reaction tube from the second reaction tube pick-up / placement position to the third reaction tube pick-up / placement position of the sample loading and discarding tray 34;
[0090] 15. Rotate the sample loading and discarding tray 34 180° to turn the reaction tube to the sample loading and liquid collection position. The sampling needle draws up the sample and enters the flow cytometry detection module 11 for detection. The detection results are displayed synchronously on the user terminal UI interface.
[0091] 16. If no retest: The sample disposal tray 34 rotates to the waste liquid extraction position, the waste liquid needle aspirates the waste liquid in the reaction tube, and then the sample disposal tray 34 rotates back to the disposal pick-up / placement position, and the waste reaction tube automatically falls into the waste bin 33; If retest: The sample disposal tray 34 rotates 180° to return the reaction tube to the third reaction tube pick-up / placement position, the gripper of the gripper arm module 6 transfers the reaction tube back to the incubation tray 30, and the gripper of the gripper arm module 6 removes the reaction tube from the incubation hole 3. 0 is transferred to the third reaction tube pick / place position of the sample loading and discarding tray 34. The sample loading and discarding tray 34 is rotated 180° to rotate the reaction tube to the sample loading and liquid collection position. The sampling needle draws the sample into the flow cytometry detection module 11 for re-inspection. The detection result is displayed synchronously on the user terminal UI interface. The sample loading and discarding tray 34 is rotated to the waste liquid collection position. The waste liquid needle draws away the waste liquid in the reaction tube. Then the sample loading and discarding tray 34 is rotated to the discard pick / place position again. The waste reaction tube automatically falls into the waste bin 33.
[0092] Example 3: Cell Detection Process
[0093] like Figure 3 As shown, taking the detection of lymphocyte subsets (TBNK) as an example, the detection process using the fully automated flow cytometer provided by this invention is as follows:
[0094] 1. First, turn on the instrument. After powering on, the instrument will perform an initialization self-test. After initialization, the instrument is ready. Place the reagents needed for the experiment into the reagent slots of the reagent tray, and place the reaction tubes (such as flow cytometer tubes) into the consumable tray module 9. Other liquids have been prepared in advance (before initialization). Then, you can select and set the corresponding execution program in the software interface.
[0095] 2. Place the sample tube into the sample rack 13. The sample entry module 1 sends the sample rack 13 to the horizontal axis 15 through the push-in area 14. During this process, the sample entry module 1 performs sample tube identification, rotation scanning and sample tube shaking.
[0096] 3. The gripper of the gripper arm module 6 takes an empty reaction tube from the consumable tray module 9 and places it into the first reaction tube pick / place position 22 of the dilution tray 21. The dilution tray 21 is rotated to move the reaction tube to the sample addition position 24.
[0097] 4. After the sample needle 28 takes a sample from the sample tube in the test tube rack 13 pushed horizontally by the horizontal movement axis 15, it adds the sample to the sample addition position 24 of the dilution tray 21.
[0098] 5. Rotate the dilution tray 21 to move the reaction tube to the reagent addition position 25;
[0099] 6. The reagent needle 18 takes fluorescent antibody reagent from the conventional reagent placement position 52 on the outer ring of the test turntable 50 and adds it to the reagent addition position 25 of the dilution tray 21;
[0100] 7. Rotate the dilution tray 21 to move the reaction tube to the oscillation mixing position 26, and oscillate the sample and reagent to mix.
[0101] 8. Rotate the dilution tray 21 to move the reaction tube to the first reaction tube pick / place position 22;
[0102] 9. The gripper of the gripper arm module 6 transfers the reaction tube to the incubation tray 30 for incubation;
[0103] 10. The gripper of the gripper arm module 6 transfers the reaction tube back from the incubation plate 30 to the first reaction tube pick-up / placement position 22 of the dilution plate 21. The dilution plate 21 rotates until the reagent addition position 25. The reagent needle 18 takes hemolysin from the hemolysin bottle placement position 17 and adds it to the reaction tube located at the reagent addition position 25. Then the dilution plate 21 rotates to the oscillation mixing position 26 to mix and then rotates to the first reaction tube pick-up / placement position 22.
[0104] 11. The gripper of the gripper arm module 6 transfers the reaction tube back from the dilution tray 21 to the incubation tray 30 for incubation;
[0105] 12. After incubation, the gripper of the gripper arm module 6 will transfer the reaction tube from the incubation tray 30 to the third reaction tube pick-up / placement position of the sample loading and discarding tray 34;
[0106] 13. Rotate the sample loading and discarding tray 34 180° to turn the reaction tube to the sample loading and liquid collection position. The sampling needle draws up the sample and enters the flow cytometry detection module 11 for detection. The detection results are displayed synchronously on the user terminal UI interface.
[0107] 14. If no retest: The sample disposal tray 34 rotates to the waste liquid extraction position, the waste liquid needle aspirates the waste liquid in the reaction tube, and then the sample disposal tray 34 rotates back to the disposal pick-up / place position, and the waste reaction tube automatically falls into the waste bin 33; If retest: The sample disposal tray 34 rotates 180° to return the reaction tube to the third reaction tube pick-up / place position, the gripper of the gripper arm module 6 transfers the reaction tube back to the incubation tray 30, and the gripper of the gripper arm module 6 removes the reaction tube from the incubation hole 3. 0 is transferred to the third reaction tube pick / place position of the sample loading and discarding tray 34. The sample loading and discarding tray 34 is rotated 180° to rotate the reaction tube to the sample loading and liquid collection position. The sampling needle draws the sample into the flow cytometry detection module 11 for re-inspection. The detection result is displayed synchronously on the user terminal UI interface. The sample loading and discarding tray 34 is rotated to the waste liquid collection position. The waste liquid needle draws away the waste liquid in the reaction tube. Then the sample loading and discarding tray 34 is rotated to the discard pick / place position again. The waste reaction tube automatically falls into the waste bin 33.
[0108] While the present invention has been disclosed above, it is not limited thereto. Any person skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention; therefore, the scope of protection of the present invention should be determined by the scope defined in the claims.
Claims
1. A fully automated flow cytometer, characterized in that, The device includes a dilution tray module, which comprises a dilution tray and six working positions. The dilution tray has multiple sockets that can rotate in place, and the sockets are used to place reaction tubes. The six working positions include a first reaction tube pick-up / placement position, a diluent addition position, a sample addition position, a reagent addition position, a vortex mixing position, and a waste liquid removal and tube discarding position. During the rotation of the dilution tray, the sockets can be rotated to the six working positions.
2. The fully automated flow cytometer as described in claim 1, characterized in that, It also includes a sample entry module, a sample addition module, a reagent needle module, and a reagent tray module; the reagent needle module includes a reagent needle, a reagent needle moving mechanism, and a needle washing pool, and the reagent needle moving mechanism drives the reagent needle to move between the reagent tray module, the dilution tray module, and the needle washing pool.
3. The fully automated flow cytometer as described in claim 2, characterized in that, The sample entry module includes a sample holder, a push-in area, and a transverse axis, which can drive the sample holder to move.
4. The fully automated flow cytometer as described in claim 3, characterized in that, The sample loading module includes a loading needle, a loading needle moving mechanism, and a loading needle cleaning mechanism. The loading needle and the loading needle moving mechanism are located above the dilution tray module and the sample entry module. The loading needle moving mechanism drives the loading needle to move between the dilution tray module and the sample entry module.
5. The fully automated flow cytometer as described in claim 4, characterized in that, The reagent tray module includes a refrigerated reagent position and a room temperature reagent position. The refrigerated reagent position includes a reagent tray and a diluent bottle placement position, while the room temperature reagent position includes a hemolysin placement position. The reagent tray is equipped with a turntable, which has a conventional reagent placement position and a microsphere reagent placement position. The conventional reagent placement position is located on the outer ring of the turntable, and the microsphere reagent placement position is located on the inner ring of the turntable. The turntable performs reciprocating rotational motion. A mixing component is also provided at the bottom of the microsphere reagent placement position.
6. The fully automated flow cytometer as described in claim 5, characterized in that, It also includes an incubation tray module, a magnetic separation module, a consumable tray module, a gripper arm assembly module, and a sample loading and discarding module; the incubation tray module includes an incubation tray with multiple incubation holes.
7. The fully automated flow cytometer as described in claim 6, characterized in that, The magnetic separation module includes a magnetic separation disk that can rotate in place. The magnetic separation disk is provided with multiple magnetic separation holes, and reaction tubes are placed in the magnetic separation holes. The magnetic separation disk is also provided with 6 working positions, including a second reaction tube pick-up / placement position, a 1mL PBS addition position, a mixing position, a magnetic adsorption position, a waste liquid extraction position, and a 100mL PBS addition position. The magnetic separation holes can rotate to the 6 working positions during the rotation of the magnetic separation disk in place.
8. The fully automated flow cytometer as described in claim 7, characterized in that, The consumables tray module is provided with multiple reaction tube storage positions; the gripper arm module includes a gripper and a gripper moving mechanism. After the reaction tube is gripped by the gripper, it is transferred between the consumables tray module, dilution tray module, incubation tray module, magnetic separation module and sample loading and discarding module through the gripper moving mechanism.
9. The fully automated flow cytometer as described in claim 8, characterized in that, The sample loading and discarding module includes a sample loading and discarding tray, which can rotate in place. The sample loading and discarding tray is provided with multiple sample loading and discarding holes, and reaction tubes are placed in the sample loading and discarding holes. The sample loading and discarding tray is also provided with four working positions, including a third reaction tube pick-up / placement position, a discard pick-up / placement position, a sample loading liquid pick-up position, and a waste liquid extraction position. During the rotation of the sample loading and discarding tray in place, the sample loading and discarding holes can rotate to the four working positions.
10. The fully automated flow cytometer as described in claim 9, characterized in that, It also includes a flow detection module and a rack module.