Magnetic bead removal method, device, and storage medium

Abstract

A magnetic bead removal method, device, and storage medium are provided. The method includes the steps of placing a liquid bag having a curved flow path on a magnetic platform, wherein a plane where the curved flow path is located is positioned above the magnetic platform and parallel to the magnetic platform; controlling a first flow rate control assembly to communicate the liquid bag with a sample container, thereby injecting a liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container into the curved flow path; flattening the liquid bag to increase a contact surface between a bottom surface of the curved flow path and the magnetic platform, and adsorbing immunomagnetic bead cells in the curved flow path onto the contact surface by the magnetic platform; and performing a collection operation to collect non-magnetic immune cells in the curved flow path into a collection container.

Description

Technical Field

[0001] This application claims the priority of a Chinese patent application with an application number of 202211181849.4 and an invention title of "Magnetic Bead Removal Method, Device, and Storage Medium", which was filed with the China National Intellectual Property Administration on September 27, 2022, and the entire content thereof is incorporated herein by reference. This application relates to the technical field of cell separation, and particularly to a magnetic bead removal method, device, and storage medium.

Background Art

[0002] In the field of biotechnology, it is common to select specific cells or bacteria with magnetic beads for culturing for experiments or treatments. Currently, even after using micron-scale magnetic beads to select immunomagnetic bead cells and removing non-magnetic immune cells by culturing immunomagnetic bead cells, the immunomagnetic bead cells still remain in the culture medium. In order to avoid adverse effects on experiments or medical treatments in the next step, it may be necessary to adsorb and remove the magnetic beads by a magnetic field. The inventors have found that in the prior art, in the process of removing immunomagnetic bead cells, due to the non-shearing characteristics of the fluid, the area of the fluid bag containing the culture medium is too large (with the development of the cell processing industry, the volume of samples in the liquid bag of the culture medium is increasing, such as sample volumes of 5L and 10L), and the flow rate of the liquid bag containing the culture medium flowing in the magnetic field is extremely uneven (see Figure 7). As a result, in some areas where the flow rate is too fast, some immunomagnetic bead cells escape, and consequently, the immunomagnetic bead cells are not completely adsorbed by the magnetic field, resulting in a decrease in the utilization rate of the magnetic field. To address the above problem, if the escape of immunomagnetic bead cells is reduced by slowing down the overall flow rate of the cell fluid, this solution has drawbacks such as a long magnetization time.

Summary of the Invention

Problems to be Solved by the Invention

[0003] The embodiments of this application provide a method, device, and storage medium for removing magnetic beads in order to solve problems in the prior art, such as the escape of immunomagnetic bead cells and low magnetic field utilization.

[0004] A method for removing magnetic beads, The steps include: placing a liquid bag having a curved channel on a magnetic platform, wherein the plane on which the curved channel exists is located above the magnetic platform and parallel to the magnetic platform; The steps include: controlling the first flow control assembly to connect the liquid bag and the sample container, thereby injecting the liquid containing magnetic bead-bound immune cells and non-magnetic immune cells from the sample container into the curved channel; The steps include: pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved channel and the magnetic platform, thereby causing the magnetic platform to adsorb the immunomagnetic bead cells in the curved channel onto the contact surface; The procedure includes the step of performing a collection operation to collect non-magnetic immune cells in the curved channel into a collection container.

[0005] A magnetic bead removal device comprising a pressing device and a controller for performing the magnetic bead removal method described above, wherein the controller is connected to the pressing device.

[0006] A computer-readable storage medium that stores computer-readable instructions that, when executed by a processor, implement the magnetic bead removal method described above. [Effects of the Invention]

[0007] The above-described magnetic bead removal method, device, and storage medium includes the steps of: placing a liquid bag having a curved channel on a magnetic platform, wherein the plane on which the curved channel exists is located above the magnetic platform and parallel to the magnetic platform; injecting a liquid containing magnetic bead-bound immune cells and non-magnetic immune cells from the sample container into the curved channel by controlling a first flow control assembly to connect the liquid bag and a sample container; pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved channel and the magnetic platform, thereby causing the magnetic platform to adsorb the immunomagnetic bead cells in the curved channel onto the contact surface; and performing a collection operation to collect the non-magnetic immune cells in the curved channel into a collection container. In this application, by designing the curved channel, the flow velocity of the liquid in the liquid bag is matched as closely as possible, the uniformity of the flow velocity is greatly improved, the adsorption capacity of the magnetic platform to immunomagnetic bead cells in a uniform flow field is sufficiently improved, and the utilization rate of the magnetic field is improved. Furthermore, by pressing the liquid bag flat to increase the contact surface between the bottom of the curved channel and the magnetic platform, the magnetic platform can more effectively adsorb the immunomagnetic bead cells in the curved channel with the increased contact surface. This prevents the immunomagnetic bead cells from escaping, improves the utilization of the magnetic field, and enables efficient and stable capture of the immunomagnetic bead cells while ensuring simplicity of operation.

[0008] Details of one or more embodiments of the present application are shown in the following drawings and description, and other features and advantages of the present application will become apparent from the specification, drawings and claims. [Brief explanation of the drawing]

[0009] To more clearly explain the technical solutions of the embodiments of this application, the drawings necessary for describing the embodiments of this application will be briefly described below. However, the drawings in the following description are only a few embodiments of this application, and it will be obvious to those skilled in the art that other drawings can be obtained based on these drawings without any creative effort. [Figure 1] This is a flowchart of a method for removing magnetic beads in one embodiment of the present invention. [Figure 2] This is a flowchart of step S30 of the magnetic bead removal method in one embodiment of the present invention. [Figure 3] This is a schematic diagram of the structure of a liquid bag in one embodiment of the present invention. [Figure 4] This is a schematic diagram of the structure of a liquid bag in another embodiment of the present invention. [Figure 5] A schematic diagram of the exploded structure of a pressing device according to one embodiment of the present invention. [Figure 6] This is a schematic diagram of the assembly structure of a pressing device according to one embodiment of the present invention. [Figure 7] This is a schematic diagram of the flow velocity distribution in a large-capacity liquid bag using conventional technology. [Figure 8] A schematic diagram of a computer device in one embodiment of the present invention. [Modes for carrying out the invention]

[0010] The technical solutions in the embodiments of this application will be described clearly and completely below with reference to the drawings of the embodiments of this application, although it is clear that the embodiments described are only a part of the embodiments of this application and not all of them. All other embodiments that a person skilled in the art could obtain without creative work based on the embodiments of this application are within the scope of protection of this application.

[0011] In one embodiment, as shown in Figure 1, a method for removing magnetic beads is provided, which includes the following steps S10 to S40. S10: A liquid bag 60 having a curved channel 61 is placed on a magnetic platform 10. The plane on which the curved channel 61 exists is located above the magnetic platform 10 and is parallel to the magnetic platform 10, where the magnetic platform 10 may be a magnetic platform made of an electromagnet or permanent magnet. For example, the magnetic platform 10 shown in Figure 5 may include a base 101 with a lift through hole 1011 and a permanent magnet magnetic block 102 or electromagnet attached to the base 101, wherein the base 101 is provided with a mounting groove and the magnetic block 102 is fixedly attached to the mounting groove. Understandably, the liquid bag 60 having the curved channel 61 is made of a non-magnetic material and is not attracted by magnetic force, and therefore is not affected by the magnetic field of the magnetic platform 10 below it.

[0012] S20: The liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container is injected into the curved channel 61 by controlling a first flow control assembly (not shown) to connect the liquid bag 60 and a sample container (not shown). Here, the first flow control assembly may include a peristaltic pump capable of providing power for the liquid to flow, and a pinch valve for opening and closing the first flow control assembly, and may also include a flow control valve for controlling the flow rate of the liquid, and sensors for monitoring the flow rate and pressure of the liquid. The sample container contains culture medium after cultivation, and the culture medium (i.e., the liquid containing magnetic bead-bound immune cells and non-magnetic immune cells in the sample container) contains immunomagnetic bead cells and non-magnetic immune cells without magnetic beads that have been incubated in culture medium and divided (during culture, magnetic beads do not divide, but cells do, so non-magnetic immune cells that have antibodies but do not have magnetic beads are obtained by division).

[0013] Furthermore, the liquid bag 60 includes a liquid inlet 62 that communicates with the sample container and a liquid outlet 63 that communicates with the collection container, and the curved channel 61 communicates between the liquid inlet 62 and the liquid outlet 63. In this embodiment, the first flow control assembly sends the culture medium in the sample container from the liquid inlet 62 to the curved channel 61, and furthermore, the magnetic platform 10 can adsorb immunomagnetic bead cells with the magnetic field of the magnetic platform 10. Non-magnetic immune cells other than immunomagnetic bead cells flow from the liquid outlet 63 of the curved channel 61 to the collection container. In this embodiment, the material of the piping connected to the curved channel 61 is a non-magnetic material such as PVC, and is therefore not affected by the magnetic field of the magnetic platform 10.

[0014] In one embodiment, the curved channel 61 includes a plurality of parallel straight pipe segments 611 and a connecting pipe segment 612 connected between two adjacent straight pipe segments 611, wherein the connecting pipe segment 612 is a curved pipe or a straight pipe that forms a preset inclination angle with the straight pipe segments 611 (the preset inclination angle is determined as needed, but for example, the preset inclination angle shown in Figure 4 is 90 degrees, i.e., the straight pipe segments 611 and the connecting pipe segment 612 are perpendicular to each other), and / or the curved channel 61 includes a spiral pipe segment. That is, in this embodiment, the curved channel 61 may be a spiral pipe segment shaped like a mosquito coil, or it may be a combination of the straight pipe segments 611 and the connecting pipe segment 612 shown in Figure 3 (where the connecting pipe segment 612 in Figure 3 is a curved pipe) and Figure 4 (where the connecting pipe segment 612 in Figure 4 is a straight pipe). Furthermore, in some embodiments, a curved channel 61 may be formed by combining straight pipe segments 611, connecting pipe segments 612, and spiral pipe segments. Understandably, in this application, all pipe segments of the curved channel are in the same plane, and this plane is parallel to the magnetic platform 10. In the above configuration, the curved channel 61 is affected by the boundary layer, the flow velocity of the liquid within it is relatively uniform, and the utilization rate of the magnetic field of the magnetic platform 10 is further increased. Understandably, the shape of the curved pipe may be set to other shapes as needed, as long as it has the effect of making the liquid flow velocity uniform and improving the utilization rate of the magnetic field. Furthermore, the maximum width of the curved channel 61 is 50 mm or less, and curved channels 61 with a width within this range are highly effective in improving the utilization rate of the magnetic field.

[0015] Furthermore, the liquid bag 60 further includes a buffer space, and the curved channel 61 communicates with the liquid outlet 63 through the buffer space. That is, the buffer space is provided to buffer the process of the liquid flowing through the curved pipe to the liquid outlet 63, and can also change the direction of liquid flow to slow down the flow velocity as needed. In one particular embodiment, the curved channel 61 is a spiral tube segment shaped like a mosquito coil, coiled with a 2 × mm liquid tube (i.e., 2 mm inner diameter, mm outer diameter; the liquid tube may be of other sizes), and a circular buffer space is provided at the center of the spiral tube segment, and the spiral tube segment communicates with the liquid outlet 63 through the buffer space.

[0016] S30: The liquid bag 60 is pressed to increase the contact surface between the bottom surface of the curved channel 61 and the magnetic platform 10, causing the magnetic platform 10 to adsorb the immunomagnetic bead cells in the curved channel 61 onto the contact surface. That is, the contact surface formed by the close contact between the liquid bag 60 and the magnetic platform 10 is enlarged, and in this way the magnetic platform 10 brings the enlarged contact surface and the immunomagnetic bead cells in the liquid (culture medium) in the liquid bag 60 into close contact, uniformly bringing the liquid into contact with the magnetic field of the magnetic platform 10, and adsorbing the immunomagnetic bead cells onto the enlarged contact surface. This ensures more sufficient magnetic adsorption in the magnetic sorting process and prevents the immunomagnetic bead cells from being unadsorbed and flowing out of the liquid bag 60 through the liquid outlet 63 along with non-magnetic immune cells. Furthermore, after the liquid bag 60 is pressed flat, the contact surface increases, which allows for the simultaneous magnetization and adsorption of magnetic beads in a large volume of cell solution during the magnetic sorting process. This enables simultaneous magnetization without controlling the flow rate, thereby achieving consistent magnetization.

[0017] S40: Perform a collection operation to collect the non-magnetic immune cells in the curved flow path 61 into a collection container (not shown). Further, in step S40, a second flow rate control assembly (not shown) provided between the collection container and the liquid bag 60 is controlled and activated to communicate the liquid bag 60 and the collection container, and the non-magnetic immune cells in the curved flow path 61 are transported to the collection container by the second flow rate control assembly. Thus, the non-magnetic immune cells are collected and used for experiments or medical treatment. That is, in this embodiment, the sample container and the collection container are both connected to the liquid bag 60. The first flow rate control assembly is provided between the sample container and the liquid bag 60, and the second flow rate control assembly is provided between the collection container and the liquid bag 60. The second flow rate control assembly may include a peristaltic pump that provides power for the liquid to flow and a pinch valve for opening and closing the second flow rate control assembly, and may also include a flow rate adjustment valve for controlling the liquid flow rate, a sensor for monitoring the liquid flow rate and pressure, etc. In this application, when only one of the first flow rate control assembly or the second flow rate control assembly is provided, the flow parameters of the liquid in the liquid bag 60, such as velocity, flow rate, pressure, etc., can be controlled by the first flow rate control assembly or / and the second flow rate control assembly. Also, both the first flow rate control assembly and the second flow rate control assembly may be provided.

[0018] In this application, by designing the curved flow path 61, the flow velocity of the liquid in the liquid bag 60 is made as consistent as possible, the uniformity of the flow velocity is greatly improved, the adsorption capacity of the magnetic platform 10 for immunomagnetic bead cells in a uniform flow field is sufficiently improved, and the utilization rate of the magnetic field is improved. Also, by pressing the liquid bag 60 flat to increase the contact surface between the bottom surface of the curved flow path 61 and the magnetic platform 10, the magnetic platform 10 can more fully adsorb the immunomagnetic bead cells in the curved flow path 61 with the increased contact surface, thereby avoiding the escape of the immunomagnetic bead cells, improving the utilization rate of the magnetic field, ensuring the simplicity of the operation, and enabling the efficient and stable capture of the immunomagnetic bead cells.

[0019] In one embodiment, as shown in FIG. 2, in step S30, the step of pressing the liquid bag 60 flat includes the following steps S301 and S302. S301: Determine whether the closing mechanism 30 of the pressing device 1 can be closed. As shown in FIGS. 5 and 6, the pressing device 1 includes a closing mechanism 30 attached to the magnetic platform 10 and a pressing mechanism 40 connected to the closing mechanism 30. When the closing mechanism 30 is closed, the liquid bag 60 is located in the accommodation space between the closing mechanism 30 and the magnetic platform 10. The closing mechanism 30 is attached to the magnetic platform 10, and the closing mechanism 30 can close or open the above accommodation space (when the liquid bag 60 disposed in the accommodation space bulges and the protruding height increases, the closing mechanism 30 may be pushed up by the liquid bag 60 and cannot be closed. If forced to close, the liquid bag 60 may burst or the immunomagnetic bead cells or anucleated immune cells therein may be damaged.). Also, when there is no interference by other external forces, the closing mechanism 30 can move downward by being pulled downward by the pressing mechanism 40, whereby the size of the accommodation space for disposing the liquid bag 60 between the magnetic platform 10 and the closing mechanism 30 can be adjusted.

[0020] S302: When the closing mechanism 30 can be closed, control the pressing mechanism 40 to move the closed closing mechanism 30 downward, and press the liquid bag 60 placed in the accommodation space between the magnetic platform 10 and the closing mechanism 30 flat. As can be understood, when the closing mechanism 30 can be closed, the pressing mechanism 40 drives the closed closing mechanism 30 to move downward, presses the liquid bag 60 uniformly and flatly, and can increase the contact surface formed by the close adhesion of the liquid bag 60 and the magnetic platform 10, so that the liquid in the liquid bag 60 passes uniformly through the contact surface and is distributed and contacted with the magnetic platform 10.

[0021] Furthermore, as shown in Figures 5 and 6, the pressing device 1 further includes a lift mechanism 20 connected to the closing mechanism 30, and after step S301, that is, after determining whether or not the closing mechanism 30 of the pressing device 1 can be closed, the following steps are further included. If the closing mechanism 30 is pushed up by the inflated liquid bag 60 and cannot be closed, the lift mechanism 20 is controlled to raise the closing mechanism 30, thereby increasing the storage space between the magnetic platform 10 and the closing mechanism 30. In this embodiment, when the liquid bag 60 is placed in the storage space, if the height of the liquid in the liquid bag 60 is high, the closing mechanism 30 is pushed up by the liquid bag 60 placed in the storage space and cannot be closed, in which case the lift mechanism 20 is controlled and activated to raise the closing mechanism 30, and in this way the distance between the magnetic platform 10 and the closing mechanism 30 increases, and the storage space increases with this increase in distance.

[0022] When the closing mechanism 30 rises to a preset closing height, it is sufficient to control the closing mechanism 30 to close it. In other words, when the closing mechanism 30 rises to a preset height, there is sufficient space between the closing mechanism 30 and the magnetic platform 10, and the closing mechanism 30 is not pushed up by the liquid bag 60, allowing it to close properly.

[0023] The pressing mechanism 40 is controlled to move the closed closing mechanism 30 downward, thereby pressing the liquid bag 60 placed in the containment space flat. That is, after the closing mechanism 30 is closed, the pressing mechanism 40 drives the closed closing mechanism 30 downward, pressing the liquid bag 60 uniformly and flatly, increasing the contact surface formed by the close contact between the liquid bag 60 and the magnetic platform 10, and ensuring that the liquid in the liquid bag 60 passes uniformly through this contact surface and is distributed and in contact with the magnetic platform 10.

[0024] In one embodiment, as shown in Figures 5 and 6, the magnetic platform 10 is provided with a lift through hole 1011, and the lift mechanism 20 includes a lifting assembly 201 provided at the top of the magnetic platform 10, a lift rod 202 whose tip passes through the lift through hole 1011 and is fixedly connected to the lifting assembly 201, and a lifting drive assembly 203 provided below the magnetic platform 10 and connected to the end of the lift rod 202 away from the lifting assembly 201, and the closing mechanism 30 includes a cover 301 rotatably connected to the lifting assembly 201. The shape of the cover 301 may be set according to the specific shape of the magnetic platform 10. Furthermore, the closing mechanism 30 further includes a handle 3011 attached to the cover 301, which allows the user to easily pull or close the cover 301. The shape of the lifting assembly 201 may be set as needed. For example, the lifting assembly 201 may be a frame structure placed on top of the magnetic platform 10 (or may consist of a number of individually provided assemblies, which may be driven by the lift rod 202 to move up and down), or it may be driven up and down by the lifting drive assembly 203. Understandably, the lifting drive assembly 203 may include a motor 2031, an air cylinder, or the like as a source of driving force to lift the lifting assembly 201. In one embodiment, as shown in Figures 5 and 6, the lifting assembly 201 includes a first lifting block 2011 provided at the first end of the magnetic platform 10, two second lifting blocks 2012 spaced apart at the second end of the magnetic platform 10 away from the first end, a limiting plate 2013 connected between the first lifting block 2011 and the second lifting blocks 2012, and a rotating shaft 2014 connected between the two second lifting blocks 2012, the cover 301 being rotatably connected to the rotating shaft 2014, and the second suction part 303 shown in Figure 5 being provided on the first lifting block 2011.Specifically, the limiting plates 2013 are two plates provided on opposite sides of the magnetic platform 10 so as to be parallel to each other. Thus, a limiting frame is obtained, enclosed by the first lifting block 2011, the two second lifting blocks 2012, and the two limiting plates 2013, which is used to prevent the liquid bag 60 from escaping from the sides of the storage space. This limiting frame (i.e., the main component of the lifting assembly 201) can be raised and lowered in sync with the cover 301.

[0025] Furthermore, the step of increasing the accommodation space between the magnetic platform 10 and the closing mechanism 30 by controlling the lift mechanism 20 to raise the closing mechanism 30 includes the step of controlling and activating the lift drive assembly 203, which drives the lift rod 202 to raise the lifting assembly 201 and the cover 301, sliding them along the lift through hole 1011, thereby increasing the accommodation space for arranging the liquid bag 60 between the magnetic platform 10 and the cover 301. That is, the closing mechanism 30 is rotatably mounted on the lifting assembly 201 and moves as the lifting assembly 201 moves up and down, while the lift drive assembly 203 can move the lifting assembly 201 upward along the lift through hole 1011 via the lift rod 202, thus moving the cover 301 upward and further increasing the accommodation space for arranging the liquid bag 60 between the magnetic platform 10 and the cover 301, and facilitating the closing of the closing cover 301.

[0026] Furthermore, as shown in Figures 5 and 6, the closing mechanism 30 further includes a first suction part 302 provided on the cover 301 and a second suction part 303 provided on the lifting assembly 201 at a position opposite to the first suction part 302, and the cover 301 is closed to the lifting assembly 201 by suction by the first suction part 302 and the second suction part 303. Here, at least one of the first suction part 302 and the second suction part 303 is a magnet, for example, both may be magnets, or one may be a magnet and the other a metal that has magnetic properties. The combination of the number of the first suction parts 302 and the second suction part 303, as well as their specific shape and size, may be set as needed, but are not limited here. Furthermore, in this application, the magnets may be electromagnets. In one particular embodiment, the second adsorption part 303 is an electromagnet and the first adsorption part 302 is a block of iron. In this case, when current is applied to the electromagnet, which is the second adsorption part 303, the magnetism of the electromagnet can be released. At this time, the electromagnet of the second adsorption part 303 and the first adsorption part 302 (block of iron) on the cover 301 are separated because they have a magnetic adsorption function, and therefore the cover 301 can be opened. On the other hand, when the current to the second adsorption part 303 is stopped, magnetism is generated in the electromagnet, which is the second adsorption part 303. At this time, if there is no interruption by an external force, and the block of iron on the cover 301 is within the magnetic attraction range of the electromagnet, the first adsorption part 302 on the cover 301 will be attracted to the second adsorption part 303, and the cover 301 will be closed.

[0027] In one embodiment, as shown in Figures 5 and 6, the lifting drive assembly 203 includes a motor 2031, a drive wheel 2032, a driven wheel 2033, a timing belt 2034, a rotating shaft 2035, a cam 2036, a mounting bearing 2037, and a carrier 2038 having a mounting hole 2039, wherein the carrier 2038 is mounted to the bottom of the magnetic platform 10, the rotating shaft 2035 is mounted in the mounting hole 2039 through the mounting bearing 2037, the cam 2036 is fixedly mounted to the rotating shaft 2035, the drive wheel 2032 is mounted to the output shaft of the motor 2031, the timing belt 2034 is stretched over the drive wheel 2032 and the driven wheel 2033, the driven wheel 2033 is fixedly mounted to the rotating shaft 2035, and the tip of the cam 2036 abuts against the lift rod 202. As shown in Figure 5, the motor 2031 may be fixed to the magnetic platform 10 by a fixing block. The cam 2036 of the rotating shaft 2035, the carrier 2038, and the mounting bearing 2037 are all provided in pairs symmetrically, and the lift rods 202 are also provided in pairs parallel to each other, with the ends of both lift rods 202 all connected to the lifting assembly 201, while the two cams 2036 each abut the bottom ends of the two lift rods 202.

[0028] Furthermore, the step of driving the lift rod 202 by the lift drive assembly 203 to raise the lifting assembly 201 and the cover 301 while sliding them along the lift through hole 1011 includes controlling and starting the motor 2031, which rotates the cam 2036 in the order of the drive wheel 2032, the timing belt 2034, the driven wheel 2033, and the rotating shaft 2035, and the rotating cam 2036 raises the lifting assembly 201 and the cover 301 while sliding them along the lift through hole 1011 via the lift rod 202. In other words, when the motor 2031 starts and rotates, it rotates the drive wheel 2032, and further rotates the driven wheel 2033 via the timing belt 2034. On the other hand, when the driven wheel 2033 rotates, the rotating shaft 2035 and the two cams 2036 attached to the rotating shaft 2035 rotate. At this time, the lift rod 202, which the cams 2036 contact, moves upward in conjunction with the rotation of the cams 2036, and further lifts the lifting assembly 201 and the cover 301 as a whole. At this time, as described in the above embodiment, if the second suction part 303 is an electromagnet, the power supply to the electromagnet which is the second suction part 303 is stopped. In this way, during the lifting process of the lifting assembly 201, the cover 301 also moves in conjunction with its rise and gradually becomes flush with the lifting assembly 201. When the iron mass of the first suction part 302 is attracted to the electromagnet of the second suction part 303, it is determined that the cover 301 has been closed.

[0029] Furthermore, as shown in Figure 5, the lifting drive assembly 203 further includes a sensing block 2042 attached to the rotating shaft 2035 and a photoelectric sensor 2043 attached to the magnetic platform 10. The photoelectric sensor 2043 detects that the cover 301 has risen to the preset height when the rotating shaft 2035 drives the sensing block 2042 to rotate it to a position facing the photoelectric sensor 2043. Here, the preset height refers to the maximum height that the lift rod 202 drives to raise the lifting assembly 201 and the cover 301. This preset height may be set according to the specific size of the liquid bag 60, and can be achieved by changing the size and shape of the cam 2036 as needed.

[0030] In one embodiment, as shown in Figure 5, the lifting drive assembly 203 further includes a bearing follower 2040 and a fixing plate 2041, wherein the bearing follower 2040 is attached to the bottom end of the lift rod 202 by the fixing plate 2041, and the cam 2036 abuts against the bearing follower 2040. That is, the bearing follower 2040 effectively transmits the lifting force due to the rotation of the cam 2036 to the lift rod 202, making the upward movement of the lift rod 202 more stable and controllable.

[0031] In one embodiment, as shown in Figures 5 and 6, the magnetic platform 10 is provided with a guide through hole 1012, and the pressing mechanism 40 includes a guide shaft 401, a spring 402, and a linear bearing 403 mounted in the guide through hole 1012, a stopper 4011 is provided at the bottom end of the guide shaft 401, the tip of the guide shaft 401 passes through the linear bearing 403 and is connected to the lifting assembly 201, the spring 402 is fitted onto the guide shaft 401 and both ends of the spring 402 abut against the linear bearing 403 and the stopper 4011, respectively. Furthermore, step S302 includes controlling the pressing mechanism 40 to move the closed closing mechanism 30 downward, which in turn controls the lifting drive assembly 203 to turn it off, thereby moving the guide shaft 401 downward along the guide through hole 1012 by the biasing force of the spring 402, and further moving the lifting assembly 201 and the cover 301 downward. That is, in this embodiment, the lifting drive assembly 203 having the cam 2036 can only drive the lift rod 202 to lift it, but does not have a downward pulling force to pull the lifting assembly 201 downward. Therefore, after the above-described pressing mechanism 40 is provided and the spring 402 is compressed, and the motor 2031 of the lifting drive assembly 203 is turned off (the motor 2031 is returned to its original position), the spring 402 resists its compressed state and applies a downward pulling force to the lifting assembly 201 via the guide shaft 401 (the spring 402 drives the guide shaft 401 to move downward along the guide through hole 1012), and further drives the lifting assembly 201 and the cover 301 to move downward.As can be understood, if one guide shaft 401, one spring 402, and one linear bearing 403 constitute one pressing assembly, the pressing mechanism 40 may include a plurality of pressing assemblies provided on the magnetic platform 10, for example, four sets of pressing assemblies provided at the four corners of a rectangular magnetic platform 10. In this way, the downward pulling force applied simultaneously by the four springs 402 of the four sets of pressing assemblies to the lifting assembly 201 causes the cover 301 to move the lifting assembly 201 downward and press the liquid bag 60 toward the magnetic platform 10 until it is flat, thereby uniformly distributing the liquid contained therein across the magnetic platform 10. In this invention, instead of a motor 2031 or the like, the spring 402 is pulled downward to flatten the liquid bag 60, which on the one hand allows the spring 402 to press the liquid bag 60 uniformly and flatly so that it can adhere tightly to the magnetic platform 10, and on the other hand, because the biasing force by the spring 402 is uniform and small, it does not press and break the cells inside the liquid bag 60.

[0032] In one embodiment, as shown in Figures 5 and 6, the magnetic sorting and pressing device 1 further includes a protector 50 attached to the bottom end of the magnetic platform 10, forming a mounting space between the protector 50 and the magnetic platform 10, and the lifting drive assembly 203 is mounted within the mounting space. That is, in this embodiment, the protector 50 can house and protect each device within the mounting space (e.g., the lifting drive assembly 203, the pressing mechanism 40, or other devices). Preferably, the protector 50 is a sheet metal part, which is more robust and reliable in structure.

[0033] It should be understood that the sequence numbers of each step in the above embodiment do not indicate the order of execution, and the execution order of each process should be determined by its function and internal logic, and do not restrict the implementation process of the embodiment of this application.

[0034] The present invention also provides a magnetic bead removal device comprising a pressing device 1 and a controller for performing the magnetic bead removal method described above, wherein the controller is connected to the pressing device 1.

[0035] Specifically, the controller is A step of placing a liquid bag having a curved channel on a magnetic platform, wherein the plane on which the curved channel exists is located above the magnetic platform and is parallel to the magnetic platform, The steps include: controlling the first flow control assembly to connect the liquid bag and the sample container, thereby injecting the liquid containing magnetic bead-bound immune cells and non-magnetic immune cells from the sample container into the curved channel; The steps include: pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved channel and the magnetic platform, thereby causing the magnetic platform to adsorb the immunomagnetic bead cells in the curved channel onto the contact surface; It is used to perform a collection operation to collect non-magnetic immune cells in the curved channel into a collection container.

[0036] In one embodiment, the liquid bag includes a liquid inlet that communicates with the sample container and a liquid outlet that communicates with the collection container, and the curved channel communicates between the liquid inlet and the liquid outlet.

[0037] In one embodiment, the curved channel includes a plurality of parallel straight pipe segments and a connecting pipe segment connected between two adjacent straight pipe segments, wherein the connecting pipe segment is a curved pipe or a straight pipe that forms a predetermined inclination angle with the straight pipe segments, and / or The curved channel includes a spiral tube segment.

[0038] In one embodiment, the maximum width of the curved channel is 50 mm or less, and / or The liquid bag further includes a buffer space, and the curved channel communicates with the liquid outlet via the buffer space.

[0039] In one embodiment, the step of pressing the liquid bag flat is performed as follows: A step of determining whether the closing mechanism of a pressing device can be closed, wherein the pressing device includes a closing mechanism attached to the magnetic platform and a pressing mechanism connected to the closing mechanism, and when the closing mechanism is closed, the liquid bag is located in the containment space between the closing mechanism and the magnetic platform. If the closing mechanism can be closed, the pressing mechanism is controlled to move the closed closing mechanism downward, and the liquid bag placed in the containment space between the magnetic platform and the closing mechanism is pressed flat.

[0040] In one embodiment, the pressing device further includes a lift mechanism connected to the closing mechanism, After the step of determining whether the closing mechanism of the pressing device can be closed, the controller further: If the closing mechanism is unable to close because it is pushed up by the inflated liquid bag, the lift mechanism is controlled to raise the closing mechanism, thereby increasing the accommodation space between the magnetic platform and the closing mechanism. The closing mechanism rises to a preset height at which it can be closed, and then the closing mechanism is controlled to close it. It is used to perform the steps of: controlling the pressing mechanism to move the closed closing mechanism downward, thereby pressing the liquid bag placed in the containment space flat.

[0041] In one embodiment, the magnetic platform is provided with a lift through hole, the lift mechanism includes a lifting assembly provided at the top of the magnetic platform, a lift rod whose tip passes through the lift through hole and is fixedly connected to the lifting assembly, and a lifting drive assembly provided below the magnetic platform and connected to the end of the lift rod away from the lifting assembly, the closing mechanism includes a cover rotatably connected to the lifting assembly, The step of increasing the accommodation space between the magnetic platform and the closing mechanism by controlling the lift mechanism to raise the closing mechanism is, The process includes controlling and activating the lifting drive assembly, and using the lifting drive assembly to drive the lift rod, thereby increasing the storage space for arranging the liquid bag between the magnetic platform and the cover, by sliding the lifting assembly and the cover along the lift through-hole.

[0042] In one embodiment, the lifting drive assembly includes a motor, drive wheels, driven wheels, a timing belt, a rotating shaft, a cam, a mounting bearing, and a carrier having mounting holes, the carrier being mounted to the bottom of the magnetic platform, the rotating shaft being mounted to the mounting holes by the mounting bearing, the cam being fixedly mounted to the rotating shaft, the drive wheels being mounted to the output shaft of the motor, the timing belt being stretched between the drive wheels and the driven wheels, the driven wheels being fixedly mounted to the rotating shaft, and the tip of the cam contacting the lift rod. The step of driving the lift rod by the lifting drive assembly to raise the lifting assembly and the cover while sliding them along the lift through hole is as follows: The process includes controlling and starting the motor, using the motor to rotate the cam in the order of the drive wheel, the timing belt, the driven wheel, and the rotating shaft, and using the rotating cam to raise the lifting assembly and the cover by sliding them along the lift through hole via the lift rod.

[0043] In one embodiment, the step of performing the collection operation to collect non-magnetic immune cells in the curved channel into a collection container is: The process includes controlling and activating a second flow control assembly provided between the collection container and the liquid bag to connect the liquid bag and the collection container, and transporting non-magnetic immune cells in the curved channel to the collection container by the second flow control assembly.

[0044] More detailed limitations of the pressing device 1 and controller of the magnetic bead removal device will not be described here, as they can be found in the limitations for the magnetic bead removal method described above. Each module of the controller described above may be implemented in whole or in part by software, hardware, or a combination thereof. Each module described above may be incorporated into the processor of a computer device in hardware form, or it may be independent of this processor, or it may be stored in the memory of the computer device in software form and invoked by the processor to perform the operations corresponding to each module described above. Understandably, the controller may be considered as one or more computer devices, as shown in Figure 8, the computer device includes a processor, memory, a network interface, and a database connected via a system bus. The processor of the computer device provides the ability to compute and control. The memory of the computer device includes a readable storage medium and internal memory. The readable storage medium stores an operating system, computer-readable instructions, and a database. The internal memory provides an environment for the operation of the operating system and computer-readable instructions in the readable storage medium. The database of the computer device is used to store the data used in the magnetic bead removal method in the above embodiment. The network interface of the computer device is connected to an external terminal via a network for communication. The computer-readable instruction, when executed by the processor, realizes a magnetic bead removal method. The readable storage medium according to this embodiment includes a non-volatile readable storage medium and a volatile readable storage medium.

[0045] In one embodiment, one or more readable storage media storing computer-readable instructions are provided, the readable storage media according to this embodiment includes a non-volatile readable storage medium and a volatile readable storage medium, the readable storage media storing computer-readable instructions, and when executed by one or more processors, the one or more processors are made to execute the magnetic bead removal method described above.

[0046] Specifically, when the computer-readable instruction is executed by one or more processors, A step of placing a liquid bag having a curved channel on a magnetic platform, wherein the plane on which the curved channel exists is located above the magnetic platform and is parallel to the magnetic platform, The steps include: controlling the first flow control assembly to connect the liquid bag and the sample container, thereby injecting the liquid containing magnetic bead-bound immune cells and non-magnetic immune cells from the sample container into the curved channel; The steps include: pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved channel and the magnetic platform, thereby causing the magnetic platform to adsorb the immunomagnetic bead cells in the curved channel onto the contact surface; The process involves having one or more processors perform a collection operation to collect non-magnetic immune cells in the curved channel into a collection container.

[0047] In one embodiment, the liquid bag includes a liquid inlet that communicates with the sample container and a liquid outlet that communicates with the collection container, and the curved channel communicates between the liquid inlet and the liquid outlet.

[0048] In one embodiment, the curved channel includes a plurality of parallel straight pipe segments and a connecting pipe segment connected between two adjacent straight pipe segments, wherein the connecting pipe segment is a curved pipe or a straight pipe that forms a predetermined inclination angle with the straight pipe segments, and / or The curved channel includes a spiral tube segment.

[0049] In one embodiment, the maximum width of the curved channel is 50 mm or less, and / or The liquid bag further includes a buffer space, and the curved channel communicates with the liquid outlet via the buffer space.

[0050] In one embodiment, the step of pressing the liquid bag flat is performed as follows: A step of determining whether the closing mechanism of a pressing device can be closed, wherein the pressing device includes a closing mechanism attached to the magnetic platform and a pressing mechanism connected to the closing mechanism, and when the closing mechanism is closed, the liquid bag is located in the containment space between the closing mechanism and the magnetic platform. If the closing mechanism can be closed, the pressing mechanism is controlled to move the closed closing mechanism downward, and the liquid bag placed in the containment space between the magnetic platform and the closing mechanism is pressed flat.

[0051] In one embodiment, the pressing device further includes a lift mechanism connected to the closing mechanism, After the step of determining whether the closing mechanism of the pressing device can be closed, the computer-readable instruction is executed by one or more processors, and further, If the closing mechanism is unable to close because it is pushed up by the inflated liquid bag, the lift mechanism is controlled to raise the closing mechanism, thereby increasing the accommodation space between the magnetic platform and the closing mechanism. The closing mechanism rises to a preset height at which it can be closed, and then the closing mechanism is controlled to close it. The process involves one or more processors performing the steps of: controlling the pressing mechanism to move the closed closing mechanism downward, thereby pressing the liquid bag placed in the containment space flat.

[0052] In one embodiment, the magnetic platform is provided with a lift through hole, the lift mechanism includes a lifting assembly provided at the top of the magnetic platform, a lift rod whose tip passes through the lift through hole and is fixedly connected to the lifting assembly, and a lifting drive assembly provided below the magnetic platform and connected to the end of the lift rod away from the lifting assembly, the closing mechanism includes a cover rotatably connected to the lifting assembly, The step of increasing the accommodation space between the magnetic platform and the closing mechanism by controlling the lift mechanism to raise the closing mechanism is, The process includes controlling and activating the lifting drive assembly, and using the lifting drive assembly to drive the lift rod, thereby increasing the storage space for arranging the liquid bag between the magnetic platform and the cover, by sliding the lifting assembly and the cover along the lift through-hole.

[0053] In one embodiment, the lifting drive assembly includes a motor, drive wheels, driven wheels, a timing belt, a rotating shaft, a cam, a mounting bearing, and a carrier having mounting holes, the carrier being mounted to the bottom of the magnetic platform, the rotating shaft being mounted to the mounting holes by the mounting bearing, the cam being fixedly mounted to the rotating shaft, the drive wheels being mounted to the output shaft of the motor, the timing belt being stretched between the drive wheels and the driven wheels, the driven wheels being fixedly mounted to the rotating shaft, and the tip of the cam contacting the lift rod. The step of driving the lift rod by the lifting drive assembly to raise the lifting assembly and the cover while sliding them along the lift through hole is as follows: The process includes controlling and starting the motor, using the motor to rotate the cam in the order of the drive wheel, the timing belt, the driven wheel, and the rotating shaft, and using the rotating cam to raise the lifting assembly and the cover by sliding them along the lift through hole via the lift rod.

[0054] In one embodiment, the step of performing a collection operation to collect non-magnetic immune cells in the curved channel into a collection container is, The process includes controlling and activating a second flow control assembly provided between the collection container and the liquid bag to connect the liquid bag and the collection container, and transporting non-magnetic immune cells in the curved channel to the collection container by the second flow control assembly.

[0055] Specific limitations on readable storage media can be found in the limitations of the magnetic bead removal method described above, and therefore will not be explained in detail here.

[0056] Those skilled in the art will understand that, when stored in a non-volatile readable storage medium or a volatile readable storage medium and executed, computer-readable instructions including the flow of each embodiment of the above methods can instruct the relevant hardware to realize all or part of the flow in each embodiment. References to memory, storage, databases, or other media used in each embodiment of the present application may include non-volatile and / or volatile memory. Non-volatile memory may include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM), or flash memory. Volatile memory may include random-access memory (RAM) or external cache memory. For example, and not limited to, RAM can be available in various forms such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), dual data rate SDRAM (DDRSDRAM), extended SDRAM (ESDRAM), synchronous link DRAM (SLDRAM), memory bus direct RAM (RDRAM), direct memory bus dynamic RAM (RDRAM), and memory bus dynamic RAM (RDRAM).

[0057] Those skilled in the art will understand that, for the sake of convenience and brevity of explanation, only the division of each of the above-mentioned functional units and modules will be given as an example; however, in actual applications, the division of the above functions can be carried out using different functional units and modules as needed, that is, the internal structure of the device can be divided into different functional units and modules to complete all or part of the above functions.

[0058] The embodiments described above are used solely to illustrate the technical solutions of the present application and are not intended to limit them. Although the present application has been described in detail with reference to the embodiments described above, those skilled in the art should understand that the technical solutions described in the embodiments described above may be modified or some of their technical features may be replaced with equivalent ones. However, such modifications or replacements should not cause the intent of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present application, and should all be included within the scope of protection of the present application. [Explanation of symbols]

[0059] 1 Pressing device, 10 Magnetic platform, 101 Base, 1011 Lift through hole, 1012 Guide through hole, 102 Magnetic block, 20 Lift mechanism, 201 Lifting assembly, 2011 First lifting block, 2012 Second lifting block, 2013 Limiting plate, 2014 Rotating shaft, 202 Lift rod, 203 Lifting drive assembly, 2031 Motor, 2032 Drive wheel, 2033 Driven wheel, 2034 Timing belt, 2035 Rotating shaft, 2036 Cam, 2037 Mounting bearing, 2038 Carrier, 2039 Mounting hole, 2040 Bearing follower, 2041 Fixing plate, 2042 Sensing block, 2043 Photoelectric sensor, 30 Closing mechanism, 301 Cover, 3011 Handle, 302 First suction part, 303 Second suction part, 40 Pressing mechanism, 401 Guide shaft, 402 Spring, 403 Linear bearing, 4011 Stopper, 50 Protector, 60 Liquid bag, 61 Curved channel, 611 Straight pipe segment, 612 Connecting pipe segment, 62 Liquid inlet, 63 Liquid outlet

Claims

1. A method for removing magnetic beads, A step of placing a liquid bag having a curved channel on a magnetic platform, wherein the plane on which the curved channel exists is located above the magnetic platform and is parallel to the magnetic platform, The steps include: controlling the first flow control assembly to connect the liquid bag and the sample container, thereby injecting the liquid containing magnetic bead-bound immune cells and non-magnetic immune cells from the sample container into the curved channel; The steps include: pressing the liquid bag flat to increase the contact surface between the bottom surface of the curved channel and the magnetic platform, thereby causing the magnetic platform to adsorb the immunomagnetic bead cells in the curved channel onto the contact surface; The steps include performing a collection operation to collect non-magnetic immune cells in the curved channel into a collection container, The step of pressing the liquid bag flat is, A step of determining whether the closing mechanism of a pressing device can be closed, wherein the pressing device includes a closing mechanism attached to the magnetic platform and a pressing mechanism connected to the closing mechanism, and when the closing mechanism is closed, the liquid bag is located in the containment space between the closing mechanism and the magnetic platform. If the closing mechanism can be closed, the step of controlling the pressing mechanism to move the closed closing mechanism downward, and pressing the liquid bag placed in the containment space between the magnetic platform and the closing mechanism flat, The pressing device further includes a lift mechanism connected to the closing mechanism, After the step of determining whether the closing mechanism of the pressing device can be closed, If the closing mechanism is unable to close because it is pushed up by the inflated liquid bag, the lift mechanism is controlled to raise the closing mechanism, thereby increasing the accommodation space between the magnetic platform and the closing mechanism. The closing mechanism rises to a preset height at which it can be closed, and then the closing mechanism is controlled to close it. A method for removing magnetic beads, further comprising the step of pressing the liquid bag placed in the containment space flat by controlling the pressing mechanism to move the closed closing mechanism downward.

2. The magnetic bead removal method according to claim 1, wherein the liquid bag includes a liquid inlet communicating with the sample container and a liquid outlet communicating with the collection container by the first flow control assembly, and the curved channel communicates between the liquid inlet and the liquid outlet.

3. The curved channel includes a plurality of parallel straight pipe segments and a connecting pipe segment connected between two adjacent straight pipe segments, wherein the connecting pipe segment is a curved pipe or a straight pipe that forms a predetermined inclination angle with the straight pipe segments, and / or The magnetic bead removal method according to claim 2, wherein the curved channel includes a spiral tube segment.

4. The maximum width of the curved channel is 50 mm or less, and / or The magnetic bead removal method according to claim 3, wherein the liquid bag further includes a buffer space, and the curved channel communicates with the liquid outlet via the buffer space.

5. The magnetic platform is provided with a lift through hole, and the lift mechanism includes a lifting assembly provided at the top of the magnetic platform, a lift rod whose tip passes through the lift through hole and is fixedly connected to the lifting assembly, and a lifting drive assembly provided below the magnetic platform and connected to the end of the lift rod away from the lifting assembly, and the closing mechanism includes a cover rotatably connected to the lifting assembly, The step of increasing the accommodation space between the magnetic platform and the closing mechanism by controlling the lift mechanism to raise the closing mechanism is, A method for removing magnetic beads according to claim 4, comprising the step of controlling and activating the lifting drive assembly, and using the lifting drive assembly to drive the lift rod, thereby increasing the accommodation space for arranging a liquid bag between the magnetic platform and the cover, by sliding the lifting assembly and the cover up along the lift through hole.

6. The lifting drive assembly includes a motor, drive wheels, driven wheels, a timing belt, a rotating shaft, a cam, a mounting bearing, and a carrier having mounting holes, the carrier being mounted to the bottom of the magnetic platform, the rotating shaft being mounted to the mounting holes by the mounting bearing, the cam being fixedly mounted to the rotating shaft, the drive wheels being mounted to the output shaft of the motor, the timing belt being stretched between the drive wheels and the driven wheels, the driven wheels being fixedly mounted to the rotating shaft, and the tip of the cam contacting the lift rod. The step of driving the lift rod by the lifting drive assembly to raise the lifting assembly and the cover while sliding them along the lift through hole is as follows: A method for removing magnetic beads according to claim 5, comprising the steps of controlling and starting the motor, rotating the cam in the order of the drive wheel, the timing belt, the driven wheel, and the rotating shaft by the motor, and raising the lifting assembly and the cover via the lift rod while sliding them along the lift through hole by the rotating cam.

7. The step of performing a collection operation to collect non-magnetic immune cells in the curved channel into a collection container is, A method for removing magnetic beads according to claim 1, comprising the steps of controlling and activating a second flow control assembly provided between the collection container and the liquid bag, connecting the liquid bag and the collection container, and transporting non-magnetic immune cells in the curved channel to the collection container by the second flow control assembly.

8. A magnetic bead removal device, A magnetic bead removal device comprising a pressing device and a controller for performing the magnetic bead removal method according to any one of claims 1 to 7, wherein the controller is connected to the pressing device.

9. A computer-readable storage medium, A computer-readable storage medium storing computer-readable instructions that, when executed by a processor, realize the magnetic bead removal method according to any one of claims 1 to 7.

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