Cell sorting system and automatic cell sorting method thereof

Abstract

The invention relates to a cell sorting system and an automatic cell sorting method thereof. The cell sorting system comprises a sorting module and a magnet matrix module. The sorting module is provided with a base column integrating a main liquid pipe and a central liquid pipe, and the main liquid pipe and the central liquid pipe are communicated through a connecting branch pipe. The magnet matrix module drives the multiple permanent magnets to rotate around the base column through the driving assembly, and a dynamic rotating magnetic field is generated in the main liquid flowing pipe. The magnetic field is configured to apply a radial magnetic force to adsorb positive cells to the outer tube wall of the main liquid flow tube, and the positive cells and negative cells form a magnetic laminar flow; negative cells which are not captured by magnetic force enter the central liquid pipe along the connecting branch pipe to complete negative sorting, then tangential magnetic force is applied to drag the positive cells to migrate downstream along the main liquid pipe, and finally positive sorting collection is achieved. By means of integrated module design and dynamic magnetic field control, continuity, automation and high reproducibility of the cell sorting process are achieved, the sorting efficiency, purity and recovery rate are remarkably improved, and good cell activity is kept.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedical engineering, and more particularly, to a cell sorting system and an automatic cell sorting method thereof. BACKGROUND

[0002] Cell sorting is a key core technology in cell biology, immunology, regenerative medicine and clinical cell therapy (such as CAR-T therapy, stem cell transplantation).

[0003] A device for cell sorting combined with fixed magnetic blocks is currently popular on the market, such as patent number CN105624035A. The device includes a magnetic cylinder with a cylindrical shape, and a mounting groove is arranged at the center of the magnetic cylinder. When in use, the operator needs to manually coil the spiral-shaped test tube pre-loaded with cell suspension and place it in the mounting groove of the magnetic cylinder.

[0004] In operation, a static magnetic field is generated inside the fixed magnetic block, which acts on the cell suspension in the spiral-shaped test tube. The magnetic bead-labeled positive cells are adsorbed on the inner wall of the test tube near the magnetic cylinder by the magnetic field. Then, the operator washes the adsorbed cells by manually squeezing or using a simple pump to push the buffer, and finally removes the test tube or magnetic block for elution to complete the sorting.

[0005] Although this device has a simple structure and low cost, it has several serious technical defects in actual application, making it difficult to meet the requirements of modern biomedical research and clinical application for efficiency, purity and stability: 1. Uncontrollable sorting process and poor reproducibility: Since the spiral-shaped test tube is manually "scattered" in the mounting groove, the relative position, fit, and tightness of the coil of the test tube and the magnetic cylinder are difficult to maintain consistent each time. This leads to differences in the magnetic field strength and gradient distribution of the pipeline each time, making the magnetic capture effect uncontrollable, and ultimately resulting in large fluctuations in the purity and recovery rate of the sorting results between batches, with extremely poor reproducibility.

[0006] 2. Low magnetic bead collection rate and limited purity: In a static magnetic field, the captured positive cell-magnetic bead complex tends to form larger "magnetic chains" or cell clusters on the tube wall. This clustering phenomenon, on the one hand, hinders the subsequent full flushing of the buffer, leading to the wrapping of negative cells, which are difficult to elute, thereby reducing the sorting purity. On the other hand, the tight clusters are difficult to completely dissociate and flush down in the final elution step, resulting in a decrease in the recovery rate of magnetic beads and positive cells.

[0007] 3. Cannot realize continuous sorting, low throughput: The device is essentially "batch" operation. After the completion of the sorting process (loading-washing-elution), the test tube must be manually removed, the product processed, and a new test tube reinstalled for the next operation. More importantly, since the magnetic field is static and continuously acting on the entire spiral pipeline, there is no mechanism for positive cells to escape the magnetic field and be continuously collected. Therefore, the device cannot realize continuous sampling and continuous discharge, and the sorting throughput is greatly limited, which cannot meet the needs of large-scale cell preparation. SUMMARY

[0008] The technical problem solved by the present application is to provide a cell sorting system with novel structure and capable of realizing continuous and automated operation, aiming at the above-mentioned defects of the prior art.

[0009] The technical solution adopted by the present application to solve its technical problem is as follows: In one aspect, the present application provides a cell sorting system, comprising a sorting module and a magnet matrix module; the sorting module comprises a cylindrical base column, a central liquid pipe vertically penetrating the center of the base column, a spiral main flow liquid pipe fixedly and spirally arranged on the outer surface of the base column, and a connecting branch pipe arranged in the base column and communicating with the central liquid pipe and the main flow liquid pipe; the main flow liquid pipe is provided for passing cell suspension; The magnet matrix module comprises a plurality of permanent magnets distributed circumferentially on the outer side of the base column, and a driving assembly for driving all the permanent magnets to rotate around the central axis of the base column to generate a dynamic rotating magnetic field in the main flow liquid pipe; Wherein, the dynamic rotating magnetic field is configured to: in the main flow liquid pipe, exert a radial magnetic force on the positive cells labeled with magnetic beads, the radial magnetic force being directed to the outer side wall of the main flow liquid pipe, so as to adsorb the positive cells to the outer side wall of the main flow liquid pipe, and make the negative cells form a magnet-induced laminar flow spatially separated from the positive cells under the action of fluid; the negative cells not captured by the magnetic force flow into the central liquid pipe along the connecting branch pipe; And exert a tangential magnetic force on the positive cells adsorbed to the outer side wall of the main flow liquid pipe, the tangential magnetic force being along the extension direction of the main flow liquid pipe, so as to drag the positive cells to migrate downstream along the main flow liquid pipe into a positive cell collection module.

[0010] The cell sorting system of the present application, wherein the outer surface of the base column is provided with a positioning groove from top to bottom for winding and mounting the main flow liquid pipe, the positioning groove being spiral-shaped.

[0011] The cell sorting system, wherein a plurality of the connecting branch pipes are arranged along the length direction of the central liquid pipe; one end of the connecting branch pipe is connected with the central liquid pipe, and the connection is a T-shaped structure; the other end of the connecting branch pipe is connected with the nearest main flow liquid pipe.

[0012] The cell sorting system, wherein a plurality of the connecting branch pipes are arranged along the length direction of the central liquid pipe; one end of the connecting branch pipe is connected with the central liquid pipe, and the connection is a T-shaped structure; the other end of the connecting branch pipe is connected with the nearest main flow liquid pipe.

[0013] The cell sorting system, wherein the upstream passage of the main flow liquid pipe is connected with the sample module through a first drainage pipe, and the downstream passage is connected with the positive cell collection module through a second drainage pipe.

[0014] The cell sorting system, wherein the first drainage pipe is provided with a first flow valve for controlling the flow rate of the cell suspension and a first sensor for monitoring whether there is liquid or bubbles.

[0015] The cell sorting system, wherein the second drainage pipe is sequentially provided with a second sensor for detecting whether the cell suspension exists in the main flow liquid pipe, a second flow valve for controlling the outflow rate of the positive cell suspension, and a first negative pressure pump for providing negative pressure to drain the positive cells along the liquid flow direction.

[0016] The cell sorting system, wherein the downstream passage of the central liquid pipe is connected with the waste liquid collection module through a third drainage pipe.

[0017] The cell sorting system, wherein the third drainage pipe is sequentially provided with a third sensor for monitoring whether the cell suspension exists in the central liquid pipe, a third flow valve for controlling the outflow rate of the negative cell suspension, and a second negative pressure pump for providing negative pressure to drain the negative cells along the liquid flow direction.

[0018] In another aspect, the application also provides an automatic cell sorting method using the cell sorting system as described above, wherein the method comprises the following steps: S1: cell separation step: passing the cell suspension into the main flow liquid pipe of the sorting module; starting the driving assembly of the magnet matrix module to drive all the permanent magnets to rotate around the base column to generate a dynamic rotating magnetic field in the main flow liquid pipe; applying a radial magnetic force of the dynamic rotating magnetic field to the positive cells marked by the magnetic beads to point to the outside wall of the main flow liquid pipe, adsorbing the positive cells to the outside wall, and making the negative cells form a spatially separated magnetic-induced laminar flow with the positive cells under the fluid push; S2: negative sorting step: After the magnetic-induced laminar flow is formed, under the action of fluid driving force, the negative cells not captured by magnetic force flow into the central liquid pipe from the connecting branch pipe and are collected; S3: magnetic dragging and migration step: By controlling the dynamic rotating magnetic field through the driving assembly, a tangential magnetic force along the extension direction of the main liquid pipe is exerted on the positive cells adsorbed to the outer wall of the main liquid pipe, so as to drag the positive cells to migrate downstream along the main liquid pipe; S4: positive selection step: When the positive cells migrate to the end of the region where the magnetic field of the base column is not mainly effective, buffer solution is introduced into the main liquid pipe, and the positive cells are flushed and collected into the positive cell collection module by using the fluid viscous force.

[0019] Compared with the prior art, the present application has the following beneficial effects: 1. In the present application, the spiral main liquid pipe is fixedly coiled on the base column to form a structure-integrated sorting module. This design ensures that the relative position of the pipeline and the magnetic field, the curvature radius and other geometric parameters are constant in each sorting, thereby ensuring the accuracy and consistency of the magnetic capture conditions.

[0020] 2. In the present application, the driving assembly drives a plurality of permanent magnets to rotate around the base column to generate a dynamic rotating magnetic field. The dynamic magnetic field can produce a weak "tumbling" or "shearing" effect on the cell clusters while capturing the cells, effectively inhibiting the formation of large magnetic chains. The dynamic magnetic field cooperates with the spiral flow path to realize "magnetic-induced laminar flow", that is, the positive cells and the negative cells are orderly separated in space on the cross section of the flow passage. This makes the subsequent negative selection very efficient and thorough in removing the negative cells, thereby greatly improving the sorting purity. At the same time, since the cells are not easy to cluster, they are more easily flushed down by the fluid in the last positive selection step, significantly improving the recovery rate of the positive cells.

[0021] 3. In the present application, the dynamic rotating magnetic field exerts a tangential magnetic force along the extension direction of the main liquid pipe on the positive cells adsorbed to the wall. This tangential force acts as a "magnetic conveyor belt" to actively and controllably drag the captured positive cells along the spiral pipeline to the downstream. Combined with continuous sample loading and product collection, the present application constructs a complete "continuous flow" sorting system. The cell suspension can continuously enter from the upstream, and the negative cells and the positive cells can be continuously collected from different outlets, which improves the sorting flux by several orders of magnitude, meeting the urgent needs of the modern cell therapy industry for large-scale and efficient cell preparation.

[0022] 4. This invention inhibits cell aggregation through the dynamic magnetic field itself; secondly, the tangential magnetic force continuously drags the adsorbed cells, preventing them from lingering and accumulating in a single location within the pipeline. Positive cells are continuously transported from the capture zone to the collection zone, significantly reducing the risk of stubborn blockages in the main liquid channel. This allows the system to operate stably and without failure for extended periods, ensuring the reliability of the sorting process.

[0023] In summary, this invention fundamentally innovates the principle and implementation of cell sorting through integrated modular design, dynamic magnetic field manipulation, and unique flow path layout, providing a powerful and landmark new tool for cell biology research and clinical cell therapy. Attached Figure Description

[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the present invention will be further described below in conjunction with the accompanying drawings and embodiments. 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: Figure 1 This is a schematic diagram of the structure of a cell sorting system according to Embodiment 1 of the present invention.

[0025] Figure 2 yes Figure 1 Cross-sectional view of a cell sorting system.

[0026] Figure 3 This is a schematic diagram of magneto-laminar flow in a cell sorting system according to Embodiment 1 of the present invention.

[0027] Figure 4 This is a schematic diagram of the magnetic dragging of a cell sorting system in Embodiment 1 of the present invention.

[0028] Figure 5 yes Figure 4 Schematic diagram of magnetic field detachment and collection in the middle cell.

[0029] Figure 6 This is the negative rinsing and collection process of a cell sorting system in Embodiment 1 of the present invention.

[0030] Figure 7 This is the positive collection process of a cell sorting system in Embodiment 1 of the present invention.

[0031] The reference numerals in the attached figures are as follows: 100 - Sorting module; 101 - Base column; 102 - Central liquid pipe; 103 - Main liquid pipe; 104 - Connecting branch pipe; 105 - Positioning groove; 200 - Magnet matrix module; 201 - Permanent magnet; 202 - Drive assembly; 300 - positive cell collection module; 301 - second drainage tube; 302 - second sensor, 303 - second flow valve; 304 - first negative pressure pump; 400 - sample module; 401 - first drainage tube; 402 - first flow valve; 403 - first sensor; 500 - waste liquid collection module; 501 - third drainage tube; 502 - third sensor, 503 - third flow valve; 504 - second negative pressure pump; 600 - master computer. DETAILED DESCRIPTION

[0032] The terms "first", "second", "third", and "fourth" and the like in the description and in the claims of the present application and the accompanying drawings are used for distinguishing between similar objects and not necessarily for describing a particular sequential or chronological order. The terms "include", "has", "has got", and "have" and any variations thereof are intended to cover a non-exclusive inclusion. For example, a process, method, system, product, or apparatus that comprises a list of steps or units are not necessarily limited to the listed steps or units, but can optionally further include additional steps or units not listed, or can optionally further include other steps or units inherent to such process, method, system, product, or apparatus.

[0033] Reference herein to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the application. The appearances of the phrase in various places in the specification are not necessarily all referring to the same embodiment, nor are they necessarily mutually exclusive or alternative embodiments. It is explicitly understood that the embodiments described herein can be combined with each other, implicitly or explicitly.

[0034] "Multiple" means two or more. "And / or", describes the association relationship of the associated objects, which means that there can be three relationships, for example, A and / or B, which can represent the three cases of A alone, A and B together, and B alone. The character " / " generally represents that the associated objects before and after are in an "or" relationship.

[0035] Furthermore, the terms "upper", "lower", "front", "back", "left", "right", "upper end", "lower end", "vertical", and the like indicating the orientation are all with reference to the attitude position of the device or equipment in normal use as described in the present scheme.

[0036] In order to make the purpose, technical scheme and advantages of the embodiments of the present application more clear, the technical scheme in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present application, not all the embodiments. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application.

[0037] 1. Example 1: A preferred embodiment of the present invention provides an automated cell sorting system, such as... Figures 1-7 As shown, the cell sorting system of the present invention mainly comprises three core components: a sorting module 100, a magnet matrix module 200, and a fluid control and collection unit. The sorting module 100 is the physical core of cell separation, the magnet matrix module 200 provides dynamic manipulation force, and the fluid control and collection unit is responsible for automatically delivering samples and buffer solutions and collecting sorted products.

[0038] Sorting module 100: In this embodiment, the sorting module 100 is the foundation for achieving high-precision separation, such as... Figure 1 and Figure 2 As shown, the core of the sorting module 100 is a cylindrical base column 101, precisely manufactured from biocompatible materials such as polycarbonate, cyclic olefin copolymers (COC), or glass, and is non-magnetic. A central liquid tube 102 is vertically inserted through the center of the base column 101, and a spiral main liquid tube 103 is fixedly coiled around the outer surface of the base column 101. A connecting branch tube 104 connecting the central liquid tube 102 and the main liquid tube 103 is provided inside the base column 101. The main liquid tube 103 allows cell suspension to pass through. This cell suspension contains positive and negative cells, and the magnetic beads have already labeled the positive cells before entering the main liquid tube 103. It is worth noting that the main liquid tube 103 can also supply buffer solution or other liquids as needed for experiments or other purposes.

[0039] In this embodiment, positive cells refer to cells that can be magnetically attracted, that is, target cells that combine with immunomagnetic beads conjugated with specific antibodies.

[0040] On the outer surface of the base column 101, a spiral positioning groove 105 is formed from top to bottom. A flexible main liquid pipe 103 (made of materials such as silicone or PVC) is precisely and tightly embedded and fixed in the positioning groove 105 to form a spiral flow channel with a fixed position.

[0041] This invention integrates the pipeline and the base column into a fixed module, ensuring that the relative position and radius of curvature of the pipeline and the external magnetic field are completely consistent during each sorting process. This guarantees constant magnetic capture conditions, eliminates batch-to-batch variations, and greatly improves the reproducibility of experimental results. The positioning groove 105 provides stable physical support and precise positioning for the main liquid pipe 103, preventing displacement or deformation under fluid pressure or equipment vibration. This ensures long-term operational stability and reliability, guarantees constant magnetic capture conditions, and fundamentally ensures high reproducibility of sorting results.

[0042] like Figure 2As shown, the central liquid tube 102 is vertically arranged and fixed on the central axis of the base column 101. At the lower end of the base column 101, a plurality of connecting branch pipes 104 are arranged inside. These connecting branch pipes 104 are sequentially distributed along the length direction (i.e. axial direction) of the central liquid tube 102, one end of which communicates with the central liquid tube 102 in a T-shaped structure, and the other end of which communicates with the nearest spiral main flow liquid tube 103; which ensures that the flow path from the inside of the main flow liquid tube to the central liquid tube is straight and free of sharp turns, greatly reducing fluid resistance. This enables the negative cells to be transferred at the minimum energy loss and the fastest speed, avoiding liquid retention and back mixing caused by poor flow path, further improving the sorting efficiency and thoroughness.

[0043] In this embodiment, the negative cells refer to cells that cannot be magnetically attracted, i.e. non-target cells that are not combined with immunomagnetic beads, which flow along the connecting branch pipe 104 to the central liquid tube 102 during the sorting process due to not being captured by the magnetic force.

[0044] By arranging the dedicated connecting branch pipe 104 inside the base column, the central liquid tube 102 and the connecting branch pipe 104 are connected in the shortest distance, and when the negative cells have been concentrated on the inside of the main flow liquid tube 103, the flow of negative cells is controlled by the flow difference between the main flow liquid tube and the central liquid tube, and by arranging independent negative pressure pumps at the outlets of the main flow liquid tube and the central liquid tube, the flow of the two fluids can be accurately controlled, forming a pressure difference. When negative sorting needs to be performed, the control system will increase the suction flow of the central liquid tube, while the input flow of the main flow liquid tube can be appropriately reduced. Under the driving of this flow difference, the negative cell liquid flow concentrated on the inside of the main flow liquid tube will naturally and spontaneously be "sucked" into the central liquid tube through the connecting branch pipe. Since the positive cells are firmly adsorbed on the outer wall of the main flow liquid tube by the stable radial magnetic force, this inward suction force will not disturb them, thereby achieving "non-destructive" removal of the negative cells and ensuring the purity of subsequent positive sorting. Therefore, the flow of negative cells will enter the central liquid tube through the branch pipe without disturbing the positive cells adsorbed on the outer wall of the main flow liquid tube by the magnetic force.

[0045] All fluid connections, including the main flow liquid tube, the central liquid tube, and the connecting branch pipe between them, are integrated inside the base column, forming a sealed fluid path completely isolated from the outside world. This completely eliminates the open operation steps that may exist in the prior art, completely eliminating the risk of pollutants such as microorganisms and dust in the air entering the cell suspension from the source. This fully closed flow path design fully meets the stringent requirements of the Good Manufacturing Practice for Pharmaceutical Products for the production process of cell therapy products, ensuring the safety and quality controllability of the final cell product, providing a solid guarantee for its clinical application, and meeting the GMP requirements for cell therapy products.

[0046] Magnet matrix module 200: Magnet matrix module 200 is the power source of the dynamic control and continuous separation of the present application, as shown in Figure 6 and Figure 7 Magnet matrix module 200 includes a turntable (not shown in the figure) on which a rotary motor as a driving assembly 202 is installed, and the rotary motor is controlled by an external host computer 600. The output shaft of the motor is fixedly connected with the center of the turntable; a plurality of high-strength permanent magnets 201 are symmetrically installed on the turntable; two permanent magnets are used in the embodiment, which can be neodymium-iron-boron magnets in the prior art. The two permanent magnets are symmetrically arranged on both sides of the base column 101, so that the permanent magnets 201 are distributed circumferentially around the base column 101.

[0047] The driving assembly 202 drives the permanent magnets 201 to rotate to generate a dynamic rotating magnetic field in the main flow pipe 103. The magnetic field not only generates a radial force to capture positive cells, but also generates a tangential force to drag the migration of positive cells. This force is like a "magnetic conveyor belt" that can actively and controllably drag the positive cells adsorbed on the pipe wall along the spiral pipe to the downstream. Combined with continuous sampling and collection in the downstream, a complete "continuous flow" system is formed. Cell suspension can continuously enter from the upstream, and negative cells and positive cells can be continuously collected from different outlets, respectively, which fundamentally solves the bottleneck problem of low throughput and also solves the defect that the static magnetic field in the prior art can only "grab" but cannot "send". In addition, by controlling the rotation speed of the driving assembly 202, the rotation frequency of the magnetic field can be accurately adjusted, so as to independently control the size of the radial capture force and the tangential drag force, to adapt to the separation requirements of different cell types and different magnetic bead labeling densities, greatly improving the flexibility and adaptability of the system.

[0048] The driving assembly 202 drives all the permanent magnets 201 to rotate around the center axis of the base column 101 to generate a dynamic rotating magnetic field in the main flow pipe 103. The dynamic rotating magnetic field is configured as follows: Figures 3-5 As shown in the figure, a radial magnetic force directed to the outer wall of the main flow pipe 103 is applied to the positive cells labeled by magnetic beads to adsorb the positive cells on the outer wall of the main flow pipe 103, and to make the negative cells form a magnet-induced laminar flow spatially separated from the positive cells under the push of the fluid; the negative cells flow into the center liquid pipe 102 along the connecting branch pipe 104; and a tangential magnetic force along the extension direction of the main flow pipe 103 is applied to the positive cells adsorbed on the outer wall of the main flow pipe 103 to drag the positive cells to migrate to the positive cell collection module 300 along the main flow pipe 103 to the downstream.

[0049] Under the action of magnetic field, the positive cells are adsorbed, and at the same time, the rotating magnetic field produces a weak "stirring" or "tumbling" effect on the positive cells, effectively inhibiting the formation of large "magnetic chains". More importantly, this design realizes "magnetic laminar flow", that is, the positive cells and the negative cells are orderly separated in space on the cross section of the flow channel. This makes the subsequent washing operation very efficient in removing the negative cells, greatly improving the sorting purity. At the same time, since the cells are not easy to aggregate, they are more easily flushed down by the fluid in the final elution step, thereby improving the recovery rate.

[0050] Fluid control and collection unit: Further, the fluid control and collection unit in the present application is an execution unit for realizing full-process automation. As shown in the figure, the unit includes a sample module 400, a positive cell collection module 300 and a waste liquid collection module 500. Figures 6-7

[0051] Upstream control: The upstream of the main liquid pipe 103 is connected to the sample module 400 through the first drainage pipe 401. A first flow valve 402 and a first sensor 403 (such as a bubble sensor or a photoelectric sensor in the prior art) are arranged on the pipeline.

[0052] The first flow valve 402 is used to control the flow rate of the cell suspension, and the first sensor 403 is used to monitor whether there is liquid or bubbles.

[0053] Positive cell collection: The downstream of the main liquid pipe 103 is connected to the positive cell collection module 300 through the second drainage pipe 301. A second sensor 302, a second flow valve 303 and a first negative pressure pump 304 are arranged in sequence along the liquid flow direction on the pipeline.

[0054] The second sensor 302, such as an optical sensor in the prior art, is used to detect whether there is cell suspension in the main liquid pipe 103; the second flow valve 303 is used to control the outflow rate of the positive cell suspension; and the first negative pressure pump 304 is used to provide negative pressure to drain the positive cells.

[0055] Waste liquid collection: The downstream of the center liquid pipe 102 is connected to the waste liquid collection module 500 through the third drainage pipe 501. A third sensor 502, a third flow valve 503 and a second negative pressure pump 504 are arranged in sequence along the liquid flow direction on the pipeline.

[0056] The third sensor 502, which can be a photoelectric sensor in the prior art, is used to monitor whether there is cell suspension in the center liquid pipe 102; the third flow valve 503 is used to control the outflow rate of the negative cell suspension; and the second negative pressure pump 504 is used to provide negative pressure to drain the negative cells.

[0057] As Figure 6 ​As shown, the cell suspension in sample module 400 is pumped into main flow tube 103 at a set flow rate. Meanwhile, the driving assembly is activated to drive the two permanent magnets 201 to rotate at a low speed (e.g. 10 RPM). Under the action of the dynamic rotating magnetic field, the magnetic-bead-labeled positive cells are adsorbed to the outer tube wall of main flow tube 103, while the negative cells flow in the inner side, forming a clear magnetic-induced laminar flow. The second negative pressure pump 504 is activated to apply negative pressure to central flow tube 102. Under the action of fluid thrust and / or negative pressure suction, the negative cells located in the inner side of main flow tube 103 are precisely drawn into central flow tube 102 through connecting branch tube 104, and finally collected into waste liquid collection module 500. The third sensor 502 monitors the turbidity of the effluent in real time, and when the turbidity decreases to the baseline level, it is determined that the negative sorting is completed.

[0058] As shown, under the action of the magnetic force dragged by the rotation of permanent magnets 201 and the flushing action of the buffer liquid, when the positive cells are dragged to the end of main flow tube 103 (out of the strong magnetic field area of the base column), the first negative pressure pump 304 is activated, and the buffer liquid is pumped from the upstream of main flow tube 103. After the rotation of permanent magnets 201 is stopped, the magnetic field binding is weakened, and the fluid viscous force will peel the positive cells from the tube wall and carry them through the second drainage tube 301 to collect into the positive cell collection module 300, completing the positive sorting. Figure 7

[0059] Through the combination of multiple sensors, flow valves and negative pressure pumps, and unified scheduling by the host computer 600, the system can automatically complete the whole process of sample loading, separation, washing and collection without human intervention, reducing the operation error and labor intensity. The above multiple sensors can also monitor the liquid state, pressure and flow rate in each pipeline in real time, providing feedback data for the central controller to achieve closed-loop control. For example, when the third sensor 502 detects that the turbidity of the waste liquid decreases to the threshold value, it can automatically determine that the negative sorting is completed, and proceed to the next step, realizing intelligent operation.

[0060] In this embodiment, the "liquid tubes" such as main flow tube 103, central flow tube 102, connecting branch tube 104, first drainage tube 401, second drainage tube 301 and third drainage tube 501 refer to flexible or rigid conduits through which cell suspensions, buffer liquids or other liquids pass. In order to facilitate the observation of fluid state and possible air bubbles, these conduits are preferably made of transparent medical-grade materials such as silicone, polyvinyl chloride (PVC), polycarbonate (PC) or cyclic olefin copolymer (COC).

[0061] Such transparent conduits are commonly used basic consumables in scientific research and production fields of chemistry, physics, biology, medical diagnosis, etc., and are widely used in blood analysis, liquid component separation, cell culture, reagent delivery and other scenarios.

[0062] 2. Example Two: ​Based on the above system, the application provides an efficient and automated cell sorting method, comprising the following steps: S1: cell separation step (magnetic-induced laminar flow formation): Place the cell suspension in the sample module, control the first flow valve, and pump the cell suspension in the sample module into the main flow tube at a set flow rate. At the same time, start the drive assembly to drive the permanent magnet to rotate at a low speed (such as 10 RPM).

[0063] Under the action of the dynamic rotating magnetic field, the positive cells labeled with magnetic beads are adsorbed to the outer wall of the main flow tube, and the negative cells flow on the inner side to form a clear magnetic-induced laminar flow. This step uses a dynamic magnetic field to suppress cell aggregation, laying a foundation for subsequent high-purity separation and solving the low-purity problem caused by aggregation in the background technology.

[0064] S2: negative sorting step (negative cell collection): When the liquid forming the magnetic-induced laminar flow flows to the connecting branch pipe, start the second negative pressure pump to apply negative pressure to the central liquid tube. Under the action of fluid thrust and / or negative pressure suction, the negative cells located on the inner side of the main flow tube are precisely drawn into the central liquid tube through the connecting branch pipe, and finally collected in the waste liquid collection module. The third sensor monitors the turbidity of the effluent in real time, and when the turbidity decreases to the baseline level, it is determined that the negative sorting is completed.

[0065] This step utilizes the synergistic effect of the T-shaped connecting branch pipe and the magnetic-induced laminar flow to achieve efficient and complete removal of negative cells, solving the problem of incomplete washing in the background technology.

[0066] S3: magnetic drag and migration step (positive cell delivery): At the same time or after negative sorting, increase the speed of the drive assembly (such as 60 RPM). The high-speed rotating magnetic field exerts a strong tangential magnetic force on the positive cells adsorbed to the wall, like a "magnetic conveyor belt", dragging the positive cells to migrate downstream along the spiral main flow tube. This is the core step to realize continuous sorting, solving the bottleneck of the background technology that cannot be continuously processed.

[0067] S4: positive sorting step (positive cell collection): When the positive cells are dragged to the end of the main flow tube (out of the strong magnetic field area of the base column), start the first negative pressure pump and pump the buffer from the upstream. After the magnetic field constraint is weakened, the fluid viscous force will peel the positive cells from the wall and carry them through the second drainage pipe to the positive cell collection module, completing the positive sorting. This step combines magnetic drag and fluid flushing to ensure high recovery of positive cells, solving the low recovery problem caused by aggregation in the background technology.

[0068] In summary, the application builds a stable, efficient and fully automatic continuous flow cell sorting platform by the synergistic design of modular integration, dynamic magnetic field manipulation and closed fluid control, and fully overcomes many defects of the prior art, and provides a powerful tool for cell research and clinical application.

[0069] The above merely describes preferred embodiments of the application and is not intended to limit the application, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the application shall be included in the protection scope of the application.

Claims

1. A cell sorting system, characterized by, The sorting module comprises a cylindrical base column, a center liquid pipe vertically penetrating the center of the base column, a helical main flow liquid pipe fixedly arranged on the outer surface of the base column, and a connecting branch pipe arranged in the base column and communicating with the center liquid pipe and the main flow liquid pipe; the main flow liquid pipe is used for passing cell suspension; The magnet matrix module comprises a plurality of permanent magnets circumferentially distributed on the outer side of the base column, and a driving assembly for driving all the permanent magnets to rotate around the central axis of the base column to generate a dynamic rotating magnetic field in the main flow liquid pipe; The dynamic rotating magnetic field is configured to exert a radial magnetic force on the magnetic-bead-labeled positive cells in the main flow liquid pipe, the radial magnetic force being directed to the outer side wall of the main flow liquid pipe, so as to adsorb the positive cells to the outer side wall of the main flow liquid pipe and make the negative cells form a spatially separated magnetic-induced laminar flow with the positive cells under the fluid pushing; the negative cells not captured by the magnetic force flow into the center liquid pipe along the connecting branch pipe; And a tangential magnetic force is exerted on the positive cells adsorbed to the outer side wall of the main flow liquid pipe, the tangential magnetic force being along the extension direction of the main flow liquid pipe, so as to drag the positive cells to migrate downstream along the main flow liquid pipe into a positive cell collection module.

2. The cell sorting system of claim 1, wherein, The outer surface of the base column is provided with a positioning groove from top to bottom for winding the main flow liquid pipe, the positioning groove being helical.

3. The cell sorting system of claim 1, wherein, The connecting branch pipe is provided with a plurality of connecting branch pipes, and the connecting branch pipes are located at the lower end of the base column.

4. The cell sorting system of claim 3, wherein, The connecting branch pipes are sequentially arranged along the length direction of the center liquid pipe; one end of the connecting branch pipe is in communication with the center liquid pipe, and the connection is a T-shaped structure; the other end of the connecting branch pipe is in communication with the nearest main flow liquid pipe.

5. The cell sorting system according to any one of claims 1 to 4, wherein, The upstream channel of the main flow liquid pipe is in communication with a sample module through a first drainage pipe, and the downstream channel is in communication with the positive cell collection module through a second drainage pipe.

6. The cell sorting system of claim 5, wherein, The first drainage pipe is provided with a first flow valve for controlling the flow rate of the cell suspension and a first sensor for monitoring whether there is liquid or bubbles.

7. The cell sorting system of claim 5, wherein, The second drainage pipe is sequentially provided with a second sensor for detecting whether the cell suspension exists in the main flow liquid pipe, a second flow valve for controlling the outflow speed of the positive cell suspension, and a first negative pressure pump for providing negative pressure to drain the positive cells along the liquid flow direction.

8. The cell sorting system of claim 5, wherein, The downstream channel of the center liquid pipe is in communication with a waste liquid collection module through a third drainage pipe.

9. The cell sorting system of claim 8, wherein, The third drainage pipe is sequentially provided with a third sensor for monitoring whether the cell suspension exists in the center liquid pipe, a third flow valve for controlling the outflow speed of the negative cell suspension, and a second negative pressure pump for providing negative pressure to drain the negative cells along the liquid flow direction.

10. An automated cell sorting method applied to the cell sorting system according to any one of claims 1 to 9, characterized in that, The method comprises the following steps: S1: cell separation step: Pass the cell suspension into the main flow liquid pipe of the sorting module; Start the driving assembly of the magnet matrix module to drive all the permanent magnets to rotate around the base column to generate a dynamic rotating magnetic field in the main flow liquid pipe; The dynamic rotating magnetic field is used to exert a radial magnetic force on the magnetic-bead-labeled positive cells, which is directed to the outer wall of the main flow liquid pipe, so that the positive cells are adsorbed to the outer wall of the main flow liquid pipe, and the negative cells are spatially separated from the positive cells under the fluid pushing force to form a magnetic-induced laminar flow; S2: negative sorting step: After the cell suspension forms a magnetic-induced laminar flow, under the action of the fluid pushing force, the negative cells not captured by the magnetic force flow into the center liquid pipe through the connecting branch pipe and are collected; S3: magnetic force dragging and migration step: The dynamic rotating magnetic field is controlled by the driving assembly, and a tangential magnetic force along the extension direction of the main flow liquid pipe is exerted on the positive cells adsorbed to the outer wall of the main flow liquid pipe, so as to drag the positive cells to migrate downstream along the main flow liquid pipe; S4: positive sorting step: When the positive cells migrate to the end of the region where the base column magnetic field mainly acts, buffer solution is introduced into the main flow liquid pipe, and the positive cells are flushed and collected into the positive cell collection module by using the fluid viscous force.

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