A cell sorting system and automated cell sorting method thereof

Abstract

The present application relates to a cell sorting system and its automatic cell sorting method, including a sorting module and a magnet matrix module. The sorting module has a base column integrated with a main flow tube and a center flow tube, and the two are connected through a connecting branch tube. The magnet matrix module drives a plurality of permanent magnets to rotate around the base column through a driving assembly, generating a dynamic rotating magnetic field in the main flow tube. The magnetic field is configured to apply a radial magnetic force to adsorb positive cells to the outer wall of the main flow tube and form a magnet-induced laminar flow with negative cells; the negative cells not captured by the magnetic force enter the center flow tube along the connecting branch tube to complete negative sorting, and secondly, a tangential magnetic force is applied to drag the positive cells to migrate downstream along the main flow tube, finally realizing positive sorting collection. Through the integrated module design and dynamic magnetic field control, the present application realizes the continuous, automatic and high reproducibility of the cell sorting process, significantly improves the sorting efficiency, purity and recovery rate, and maintains good cell activity.

Description

Technical Field

[0001] This invention relates to the technical field of biomedical engineering, and more specifically, to a cell sorting system and an automated cell sorting method thereof. Background Technology

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

[0003] Currently, a device that combines a fixed magnetic block for cell sorting is popular on the market, such as patent number CN105624035A. This device includes a cylindrical magnetic column with a mounting groove in the center. In use, the operator needs to manually coil a spiral test tube pre-filled with cell suspension and place it in the mounting groove of the magnetic column.

[0004] During operation, a static magnetic field is generated inside the fixed magnetic block, which acts on the cell suspension in the spiral test tube. Positive cells labeled with magnetic beads are adsorbed onto the inner wall of the test tube near the magnetic column by the magnetic field. The operator then washes the adsorbed cells by manually squeezing or using a simple pump to push the buffer solution. Finally, the test tube or magnetic block is removed for elution, completing the sorting process.

[0005] Although this device is simple in structure and low in cost, it has revealed several serious technical defects in practical applications, making it difficult to meet the requirements of modern biomedical research and clinical applications for efficiency, purity, and stability:

[0006] 1. Uncontrollable sorting process and poor reproducibility: Because the spiral test tubes are manually "scattered" in the mounting slot, it is difficult to maintain consistency in the relative position, fit, and tightness of the coiling between the test tubes and the magnetic column each time. This results in differences in the magnetic field strength and gradient distribution of the pipeline during each sorting, making the magnetic capture effect uncontrollable. Ultimately, this leads to large batch-to-batch fluctuations in the purity and recovery rate of the sorting results, resulting in extremely poor reproducibility.

[0007] 2. Low magnetic bead collection rate and limited purity: In a static magnetic field, the captured positive cell-magnetic bead complex tends to aggregate on the tube wall, forming large "magnetic chains" or cell clusters. This aggregation hinders thorough rinsing with buffer solution, causing negative cells to be encapsulated and difficult to elute, thus reducing sorting purity. Furthermore, the tightly packed clusters are difficult to completely disintegrate and wash off in the final elution step, resulting in a reduced recovery rate of both magnetic beads and positive cells.

[0008] 3. Inability to achieve continuous sorting and low throughput: This device is essentially a "batch" operation. After one sorting cycle (sample loading-washing-elution), the test tube must be manually removed, the product processed, and a new test tube reinstalled for the next round. More importantly, because the magnetic field is static and continuously acts on the entire spiral tube, there is no mechanism to allow positive cells to be removed from the magnetic field and continuously collected. Therefore, this device cannot achieve continuous sample loading and continuous product discharge, and the sorting throughput is greatly limited, failing to meet the needs of large-scale cell preparation. Summary of the Invention

[0009] The technical problem this invention aims to solve is to provide a novel cell sorting system capable of continuous and automated operation, addressing the aforementioned deficiencies of existing technologies. This system significantly improves cell sorting efficiency and purity through the synergistic effect of an innovative dynamic rotating magnetic field and a helical flow path, while better preserving cell viability.

[0010] The technical solution adopted by this invention to solve its technical problem is as follows:

[0011] On one hand, the present invention provides a cell sorting system, comprising a sorting module and a magnet matrix module; the sorting module includes a cylindrical base column, a central liquid tube vertically passing through the center of the base column, a spiral main liquid tube fixedly coiled around the outer surface of the base column, and a connecting branch pipe connecting the central liquid tube and the main liquid tube inside the base column; the main liquid tube allows cell suspension to pass through;

[0012] The magnet matrix module includes multiple permanent magnets circumferentially distributed on the outside of the base column, and a drive component for driving all the permanent magnets to rotate about the central axis of the base column to generate a dynamic rotating magnetic field in the main fluid pipe.

[0013] The dynamic rotating magnetic field is configured such that: within the main fluid tube, a radial magnetic force is applied to the positive cells labeled with magnetic beads, pointing towards the outer wall of the main fluid tube, so as to adsorb the positive cells onto the outer wall of the main fluid tube, and cause the negative cells to form a spatially separated magneto-laminar flow with the positive cells under the propulsion of the fluid; the negative cells not captured by the magnetic force flow into the central fluid tube along the connecting branch tube;

[0014] A tangential magnetic force is applied to the positive cells adsorbed on the outer wall of the main fluid tube along the extension direction of the main fluid tube to drag the positive cells downstream along the main fluid tube to the positive cell collection module.

[0015] In the cell sorting system of the present invention, the outer surface of the base column is provided with a positioning groove from top to bottom for winding and installing the main fluid tube, and the positioning groove is spiral-shaped.

[0016] In the cell sorting system of the present invention, multiple connecting branches are provided, and all of the multiple connecting branches are located at the lower end of the base column.

[0017] In the cell sorting system of the present invention, a plurality of connecting branches are arranged sequentially along the length of the central liquid tube; one end of each connecting branch is connected to the central liquid tube, and the connection point is a T-shaped structure; the other end of each connecting branch is connected to the nearest main liquid tube.

[0018] In the cell sorting system of the present invention, the upstream channel of the main fluid tube is connected to the sample module through a first drainage tube, and the downstream channel is connected to the positive cell collection module through a second drainage tube.

[0019] The cell sorting system of the present invention includes a first flow valve for controlling the flow rate of the cell suspension and a first sensor for monitoring the presence or absence of liquid or air bubbles on the first drainage tube.

[0020] The cell sorting system of the present invention includes, in the second drainage tube, a second sensor for detecting whether the cell suspension exists in the main liquid tube, 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, arranged sequentially along the liquid flow direction.

[0021] In the cell sorting system of the present invention, the downstream channel of the central liquid tube is connected to the waste liquid collection module through a third drainage tube.

[0022] The cell sorting system of the present invention includes, in the third drainage tube, a third sensor for monitoring the presence of cell suspension in the central liquid tube, a third flow valve for controlling the outflow rate of negative cell suspension, and a second negative pressure pump for providing negative pressure to drain negative cells, arranged sequentially along the liquid flow direction.

[0023] On the other hand, the present invention also provides an automated cell sorting method, using any of the cell sorting systems described above, wherein the method includes the following steps:

[0024] S1: Cell separation steps:

[0025] The cell suspension is fed into the main liquid tube of the sorting module;

[0026] The drive component of the magnet matrix module is activated, driving all permanent magnets to rotate around the base column to generate a dynamic rotating magnetic field within the main fluid tube.

[0027] The dynamic rotating magnetic field is used to apply a radial magnetic force to the positive cells labeled with magnetic beads, pointing towards the outer wall of the main fluid tube, so that the positive cells are adsorbed onto the outer wall of the tube, and the negative cells are spatially separated from the positive cells by the fluid.

[0028] S2: Negative sorting steps:

[0029] After the cell suspension forms a magneto-laminar flow, under the action of fluid propulsion, negative cells that are not captured by the magnetic force flow along the connecting branch tube into the central liquid tube and are collected.

[0030] S3: Magnetic Driving and Migration Steps:

[0031] The dynamic rotating magnetic field is controlled by the driving component to apply a tangential magnetic force along the extension direction of the main fluid tube to the positive cells adsorbed on the outer wall of the main fluid tube, so as to drag the positive cells downstream along the main fluid tube.

[0032] S4: Positive sorting steps:

[0033] When positive cells migrate to the end of the column where they are no longer in the main area of ​​magnetic field, buffer solution is introduced into the main flow channel to flush and collect the positive cells into the positive cell collection module using fluid viscosity.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] 1. This invention forms a structurally integrated sorting module by fixing the spiral main liquid pipe around a base column. This design ensures that the relative position of the pipe and the magnetic field, the radius of curvature, and other geometric parameters remain constant in each sorting, thereby guaranteeing the accuracy and consistency of the magnetic capture conditions.

[0036] 2. This invention drives multiple permanent magnets to rotate around a base column via a driving component, generating a dynamic rotating magnetic field. This dynamic magnetic field, while capturing cells, also produces a weak "tumbling" or "shearing" effect on cell clusters, effectively suppressing the formation of large magnetic chains. The dynamic magnetic field, in conjunction with the helical flow path, achieves "magnetically induced laminar flow," meaning that positive and negative cells are spatially separated in an orderly manner across the flow channel cross-section. This allows subsequent negative sorting to remove negative cells very efficiently and thoroughly, thus greatly improving sorting purity. Simultaneously, because cells are less prone to clustering, they are more easily flushed off by the fluid in the final positive sorting step, significantly improving the recovery rate of positive cells.

[0037] 3. This invention utilizes a dynamic rotating magnetic field to apply a tangential magnetic force along the direction of the main fluid flow to positive cells adsorbed on the tube wall. This tangential force acts like a "magnetic conveyor belt," actively and controllably dragging the captured positive cells continuously downstream along the spiral tube. Combined with continuous sample loading and product collection, this invention constructs a complete "continuous flow" sorting system. Cell suspension can continuously enter from upstream, while negative and positive cells can be continuously collected from different outlets, increasing sorting throughput by several orders of magnitude and meeting the urgent needs of the modern cell therapy industry for large-scale, efficient cell preparation.

[0038] 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.

[0039] 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

[0040] 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:

[0041] Figure 1 This is a schematic diagram of the structure of a cell sorting system according to Embodiment 1 of the present invention.

[0042] Figure 2 yes Figure 1 Cross-sectional view of a cell sorting system.

[0043] Figure 3 This is a schematic diagram of magneto-laminar flow in a cell sorting system according to Embodiment 1 of the present invention.

[0044] Figure 4 This is a schematic diagram of the magnetic dragging of a cell sorting system in Embodiment 1 of the present invention.

[0045] Figure 5 yes Figure 4 Schematic diagram of magnetic field detachment and collection in the middle cell.

[0046] Figure 6 This is the negative rinsing and collection process of a cell sorting system in Embodiment 1 of the present invention.

[0047] Figure 7 This is the positive collection process of a cell sorting system in Embodiment 1 of the present invention.

[0048] The accompanying figure is labeled as follows:

[0049] 100 - Sorting module; 101 - Base column; 102 - Central liquid pipe; 103 - Main liquid pipe; 104 - Connecting branch pipe; 105 - Positioning groove;

[0050] 200 - Magnet matrix module; 201 - Permanent magnet; 202 - Drive assembly;

[0051] 300 - Positive cell collection module; 301 - Second drainage tube; 302 - Second sensor; 303 - Second flow valve; 304 - First negative pressure pump;

[0052] 400 - Sample module; 401 - First drainage tube; 402 - First flow valve; 403 - First sensor;

[0053] 500 - Waste liquid collection module; 501 - Third drainage tube; 502 - Third sensor; 503 - Third flow valve; 504 - Second negative pressure pump;

[0054] 600 - Main control computer. Detailed Implementation

[0055] The terms "first," "second," "third," and "fourth," etc., used in the specification, claims, and accompanying drawings of this invention are used to distinguish different objects, not to describe a specific order. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. For example, a process, method, system, product, or apparatus that includes a series of steps or units is not limited to the listed steps or units, but may optionally include steps or units not listed, or may optionally include other steps or units inherent to these processes, methods, products, or apparatuses.

[0056] In this document, the term "embodiment" means that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of the invention. The appearance of this phrase in various places throughout the specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment mutually exclusive with other embodiments. It will be explicitly and implicitly understood by those skilled in the art that the embodiments described herein can be combined with other embodiments.

[0057] "Multiple" refers to two or more. "And / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A alone, A and B simultaneously, or B alone. The character " / " generally indicates that the preceding and following related objects have an "or" relationship.

[0058] Furthermore, the terms indicating orientation, such as "up, down, front, back, left, right, upper end, lower end, longitudinal," etc., are all based on the posture and position of the device or equipment described in this solution during normal use.

[0059] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, a clear and complete description will be provided below in conjunction with the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the protection scope of the present invention.

[0060] 1. Example 1:

[0061] A preferred embodiment of the present invention provides an automated cell sorting system, such as... Figure 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.

[0062] Sorting module 100:

[0063] 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.

[0064] 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.

[0065] 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.

[0066] 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.

[0067] like Figure 2 As shown, the central liquid tube 102 is vertically inserted and fixed on the central axis of the base column 101. At the lower end of the base column 101, multiple connecting branch tubes 104 are internally arranged. These connecting branch tubes 104 are distributed sequentially along the length (i.e., axial direction) of the central liquid tube 102, with one end connected to the central liquid tube 102 in a T-shaped structure, and the other end connected to the nearest spiral main liquid tube 103; this ensures a straight flow path from the inside of the main liquid tube to the central liquid tube without sharp turns, greatly reducing fluid resistance. This allows negative cells to be transferred with minimal energy loss and at the fastest speed, avoiding liquid retention and backmixing caused by obstructed flow paths, further improving sorting efficiency and thoroughness.

[0068] In this embodiment, negative cells refer to cells that cannot be attracted by magnets, i.e., non-target cells that do not bind to immunomagnetic beads. During the sorting process, they flow along the connecting branch tube 104 toward the central liquid tube 102 because they are not captured by magnets.

[0069] By installing a dedicated connecting branch pipe 104 inside the column, the central fluid pipe 102 and the connecting branch pipe 104 are connected by the shortest distance. When negative cells are concentrated inside the main fluid pipe 103, the flow of negative cell fluid is controlled by the flow difference between the main fluid pipe and the central fluid pipe. By installing independent negative pressure pumps at the outlets of the main fluid pipe and the central fluid pipe, the flow rates of the two fluids can be precisely controlled to form a pressure difference. When negative sorting is required, the control system increases the suction flow rate of the central fluid pipe and appropriately reduces the input flow rate of the main fluid pipe. Driven by this flow difference, the negative cell fluid concentrated inside the main fluid pipe will naturally and smoothly be "siphoned" into the central fluid pipe through the connecting branch pipe. Because positive cells are firmly adsorbed onto the outer wall of the main liquid tube by a stable radial magnetic force, the suction force from the inside out will not disturb them, thus achieving the "non-destructive" removal of 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 tube without disturbing the positive cells that are adsorbed onto the outer wall of the main liquid tube by the magnetic force.

[0070] All fluid connections, including the main fluid line, the central fluid line, and the connecting branches between them, are integrated within the column, forming a completely closed fluid path isolated from the outside environment. This completely eliminates any open operation steps that may exist in existing technologies, preventing the risk of airborne microorganisms, dust, and other contaminants entering the cell suspension at the source. This fully enclosed flow path design fully complies with the stringent requirements of Good Manufacturing Practices (GMP) for cell therapy product manufacturing processes, ensuring the safety and quality control of the final cell product, providing a solid guarantee for its clinical application, and meeting GMP requirements for cell therapy products.

[0071] Magnet Matrix Module 200:

[0072] The magnet matrix module 200 is the power source for dynamic control and continuous sorting in this invention, such as... Figure 6 and Figure 7 As shown, the magnet matrix module 200 includes a turntable (not shown) on which a rotary motor, serving as a drive component 202, is mounted. The rotary motor is controlled by an external main control computer 600. The output shaft of the motor is fixedly connected to the center of the turntable. Multiple high-strength permanent magnets 201 are symmetrically mounted on the turntable. In this embodiment, two permanent magnets are used, which can be neodymium iron boron magnets as used 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 circumferentially distributed around the base column 101.

[0073] The drive component 202 drives the permanent magnet 201 to rotate, generating a dynamic rotating magnetic field within the main liquid tube 103. This magnetic field not only generates a radial force to capture positive cells but also a tangential force to drag positive cells downstream. This force acts like a "magnetic conveyor belt," actively and controllably dragging positive cells adsorbed on the tube wall downstream along the spiral path. Combined with continuous sample introduction and downstream collection, a complete "continuous flow" system is formed. Cell suspension can continuously enter from upstream, while negative and positive cells can be continuously collected from different outlets, fundamentally solving the bottleneck problem of low throughput and overcoming the limitation of static magnetic fields in existing technologies that can only "grab" but not "deliver." Furthermore, by controlling the rotational speed of the drive component 202, the rotation frequency of the magnetic field can be precisely adjusted, thereby independently controlling the magnitude of the radial capture force and tangential dragging force to adapt to the sorting requirements of different cell types and different magnetic bead labeling densities, greatly improving the system's flexibility and adaptability.

[0074] The drive assembly 202 drives all permanent magnets 201 to rotate around the central axis of the base column 101, thereby generating a dynamic rotating magnetic field within the main fluid pipe 103. The dynamic rotating magnetic field is configured as follows: Figure 3-5 As shown, within the main liquid tube 103, a radial magnetic force is applied to the positive cells labeled with magnetic beads, pointing towards the outer wall of the main liquid tube 103, so as to adsorb the positive cells onto the outer wall of the main liquid tube 103, and to cause the negative cells to form a spatially separated magneto-laminar flow with the positive cells under the propulsion of the fluid; the negative cells flow into the central liquid tube 102 along the connecting branch tube 104; and a tangential magnetic force is applied to the positive cells adsorbed on the outer wall of the main liquid tube 103 along the extension direction of the main liquid tube 103, so as to drag the positive cells downstream along the main liquid tube 103 to the positive cell collection module 300.

[0075] Under the influence of the magnetic field, positive cells are adsorbed, and the rotating magnetic field creates a weak "stirring" or "tumbling" effect, effectively suppressing the formation of large "magnetic chains." More importantly, this design achieves "magnetically induced laminar flow," meaning that positive and negative cells are spatially separated in an orderly manner across the flow channel cross-section. This allows subsequent washing operations to remove negative cells very efficiently, greatly improving sorting purity. Simultaneously, because the cells are less prone to clumping, they are more easily washed away by the fluid in the final elution step, thus increasing the recovery rate.

[0076] Fluid control and collection unit:

[0077] Furthermore, the fluid control and collection unit in this invention is an execution unit that realizes full-process automation. For example... Figure 6-7As shown, the unit includes a sample module 400, a positive cell collection module 300, and a waste liquid collection module 500.

[0078] Upstream control: The upstream of the main liquid pipe 103 is connected to the sample module 400 through the first drainage pipe 401. The pipeline is equipped with a first flow valve 402 and a first sensor 403 (such as a bubble sensor or photoelectric sensor in the prior art).

[0079] 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 the presence of liquid or air bubbles.

[0080] Positive cell collection: The downstream of the main fluid pipe 103 is connected to the positive cell collection module 300 via the second drainage pipe 301. A second sensor 302, a second flow valve 303, and a first negative pressure pump 304 are sequentially installed along the liquid flow direction on the pipeline.

[0081] 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 fluid tube 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.

[0082] Waste liquid collection: Downstream of the central liquid pipe 102, it is connected to the waste liquid collection module 500 via the third drainage pipe 501. Along the liquid flow direction, the pipeline is equipped with a third sensor 502, a third flow valve 503, and a second negative pressure pump 504 in sequence.

[0083] The third sensor 502 can be a photoelectric sensor in the prior art, used to monitor whether there is cell suspension in the central liquid tube 102; the third flow valve 503 is used to control the outflow rate of negative cell suspension; the second negative pressure pump 504 is used to provide negative pressure to drain negative cells.

[0084] like Figure 6 As shown, the cell suspension in sample module 400 is pumped into the main liquid tube 103 at a set flow rate. Simultaneously, the drive assembly is activated, driving two permanent magnets 201 to rotate at a low speed (e.g., 10 RPM). Under the influence of the dynamic rotating magnetic field, positive cells labeled with magnetic beads are adsorbed onto the outer wall of the main liquid tube 103, while negative cells flow on the inner side, forming a clear magneto-laminar flow. The second negative pressure pump 504 is activated, applying negative pressure to the central liquid tube 102. Under the action of fluid thrust and / or negative pressure suction, negative cells located inside the main liquid tube 103 are precisely drawn into the central liquid tube 102 through the connecting branch tube 104, and finally collected in the waste liquid collection module 500. The third sensor 502 monitors the turbidity of the effluent in real time; when the turbidity drops to the baseline level, negative sorting is considered complete.

[0085] like Figure 7As shown, positive cells are dragged to the end of the main flow tube 103 (away from the strong magnetic field region of the base column) by the magnetic force generated by the rotation of the permanent magnet 201 and the rinsing effect of the buffer solution. At this point, the first negative pressure pump 304 is activated, and buffer solution is pumped in from the upstream of the main flow tube 103. After the permanent magnet 201 stops rotating, causing the magnetic field binding to weaken, the fluid viscosity force peels the positive cells off the tube wall and carries them through the second drainage tube 301 to the positive cell collection module 300, completing the positive sorting.

[0086] By combining multiple sensors, flow valves, and negative pressure pumps, and under the unified control of the main control computer 600, the system can automatically complete the entire process of sample loading, separation, washing, and collection without manual intervention, reducing operational errors and labor intensity. These multiple sensors can also monitor the liquid state, pressure, and flow rate in each pipeline in real time, providing feedback data to the central controller for closed-loop control. For example, when the third sensor 502 detects that the turbidity of the waste liquid has dropped to a threshold, it can automatically determine that negative separation is complete and proceed to the next step, achieving intelligent operation.

[0087] In this embodiment, the "liquid tubes" such as the main fluid tube 103, central fluid 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 solutions, or other liquids pass. To facilitate observation of the fluid state and potential air bubbles, these conduits are preferably made of transparent medical-grade materials, such as silicone, polyvinyl chloride (PVC), polycarbonate (PC), or cyclic olefin copolymers (COC).

[0088] These transparent catheters are basic consumables commonly used in scientific research and production fields such as chemistry, physics, biology, and medical diagnostics, and are widely used in various scenarios such as blood analysis, liquid component separation, cell culture, and reagent delivery.

[0089] 2. Example 2:

[0090] Based on the above system, the present invention provides an efficient and automated cell sorting method, comprising the following steps:

[0091] S1: Cell separation step (forming magneto-laminar flow):

[0092] The cell suspension is placed in the sample module, and the first flow valve is controlled to pump the cell suspension from the sample module into the main fluid tube at a set flow rate. At the same time, the drive assembly is activated to drive the permanent magnet to rotate at a low speed (e.g., 10 RPM).

[0093] Under the influence of a dynamic rotating magnetic field, positively labeled cells are adsorbed onto the outer wall of the main liquid tube, while negatively labeled cells flow on the inner side, forming a clear magneto-laminar flow. This step utilizes the dynamic magnetic field to suppress cell aggregation, laying the foundation for subsequent high-purity separation and solving the problem of low purity caused by aggregation in the previous technique.

[0094] S2: Negative sorting step (negative cell collection):

[0095] When the magneto-laminar fluid reaches the connecting branch pipe, the second negative pressure pump is activated to apply negative pressure to the central liquid pipe. Under the action of fluid thrust and / or negative pressure suction, negative cells located inside the main liquid pipe are precisely drawn into the central liquid pipe 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 drops to the baseline level, the negative sorting is considered complete.

[0096] This step utilizes the synergistic effect of T-shaped connecting branches and magneto-laminar flow to achieve efficient and thorough removal of negative cells, solving the problem of incomplete washing in the previous technique.

[0097] S3: Magnetic Driving and Migration Steps (Positive Cell Delivery):

[0098] During or after negative sorting, the rotational speed of the drive assembly is increased (e.g., to 60 RPM). The high-speed rotating magnetic field applies a strong tangential magnetic force to the positive cells adsorbed on the tube wall, dragging them downstream along the spiral main liquid tube like a "magnetic conveyor belt." This is the core step in achieving continuous sorting, overcoming the bottleneck of previous technologies that could not process continuously.

[0099] S4: Positive cell sorting step (positive cell collection):

[0100] When positive cells are dragged to the end of the main liquid tube (away from the strong magnetic field zone of the column), the first negative pressure pump is activated, pumping buffer solution upstream. As the magnetic field binding weakens, fluid viscosity detaches the positive cells from the tube wall and carries them through the second drainage tube to the positive cell collection module, completing the positive sorting process. This step combines magnetic dragging and fluid rinsing, ensuring a high recovery rate of positive cells and solving the problem of low recovery rates caused by aggregation in the prior art.

[0101] In summary, this invention constructs a stable, efficient, and fully automated continuous flow cell sorting platform through the synergistic design of modular integration, dynamic magnetic field manipulation, and closed-loop fluid control. It comprehensively overcomes many shortcomings of existing technologies and provides a powerful tool for cell research and clinical applications.

[0102] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

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 connected 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; the connecting branch pipe is located at the lower end of the base column, one end of the connecting branch pipe is connected with the center liquid pipe, and the connection is a T-shaped structure.

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 and installing the main flow liquid pipe, and the positioning groove is helical.

3. The cell sorting system of claim 1, wherein, The connecting branch pipe is provided with a plurality of.

4. The cell sorting system of claim 3, wherein, The plurality of connecting branch pipes are sequentially arranged along the length direction of the center liquid pipe; the other end of the connecting branch pipe is connected 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 connected with a sample module through a first drainage pipe, and the downstream channel is connected 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 connected 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 the 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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