Pipeline system for cell sorting, cell sorting equipment and control method

Abstract

The invention discloses a pipeline system for cell sorting, cell sorting equipment and a control method. The pipeline system comprises a magnetic field assembly and a plurality of sorting containers arranged in the magnetic field assembly, wherein the sorting containers are arranged in parallel.

Description

A pipeline system for cell sorting, cell sorting equipment and control method

[0001] Priority information

[0002] This invention claims priority to Chinese patent application No. 202210667841.2 filed on June 14, 2022, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This specification relates to the field of cell sorting technology, and in particular to a piping system, cell sorting equipment, and control method for cell sorting. Background Art

[0004] With the continuous development of cell therapy and gene therapy technologies, cell-related research has gradually become a research hotspot in current medicine and even the entire life science field. The acquisition of target cells is a key step in cell therapy, and to obtain relatively pure target cells, cell sorting and purification have become key issues. Cell sorting is the separation of a single cell from a multi-cell sample. Specifically, cells are labeled with antibodies. The antibodies used are fluorescently labeled or linked to immunomagnetic beads, and the properties of fluorescence or immunomagnetic beads are used to separate cells.

[0005] Immunomagnetic cell sorting is a simple process with low technical requirements. The sorted cells are highly sensitive, pure, and have good cell viability and recovery rates, with minimal impact on downstream applications and broad application prospects. Existing immunomagnetic cell sorting methods primarily include manual magnetic column sorting and automated sorting instruments. However, existing immunomagnetic cell sorting equipment has limited single-sample sorting volume, long sorting times, and significant impacts on cell viability and normal cell function.

[0006] Therefore, it is necessary to provide a pipeline system, cell sorting equipment and control method for cell sorting to achieve high-quality cell sorting with large volume, high purity, high yield, reproducibility and normal cell viability and cell function in a short time.

[0007] Summary of the Invention

[0008] An embodiment of the present specification provides a piping system for cell sorting, the piping system comprising: a magnetic field assembly and a plurality of sorting containers disposed in the magnetic field assembly; wherein the plurality of sorting containers are disposed in parallel.

[0009] In some embodiments, the piping system further includes a sample bag, and parallel piping disposed between the sample bag and the plurality of sorting containers.

[0010] In some embodiments, the magnetic field assembly includes a plurality of magnetic field modules, wherein each of the plurality of sorting containers is disposed in one of the magnetic field modules.

[0011] In some embodiments, the magnetic field assembly includes a magnetic field module, wherein the multiple sorting containers share one magnetic field module.

[0012] In some embodiments, the magnetic field assembly includes a plurality of magnetic field modules, wherein the number of the sorting containers is greater than the number of the magnetic field modules, and at least two of the sorting containers share one magnetic field module.

[0013] In some embodiments, the pipeline system further includes a coarse sorting container, which is arranged in series before the multiple sorting containers.

[0014] In some embodiments, the pipeline system further includes a cell transfer bag, a first pipeline, and a second pipeline; wherein, the two ends of the first pipeline are respectively connected to the outlet end of the coarse sorting container and the first interface of the cell transfer bag, and the second pipeline is arranged between the second interface of the cell transfer bag and the inlet ends of the multiple sorting containers.

[0015] In some embodiments, the pipeline system further includes a first bubble sensor disposed on the second pipeline.

[0016] In some embodiments, the pipeline system includes a first parallel pipeline, a second parallel pipeline and a junction pipeline; wherein, one end of the first parallel pipeline and the second parallel pipeline is connected to the outlet end of the coarse sorting container, and the other end is connected to the junction pipeline; at least one sorting container is respectively provided on the first parallel pipeline and the second parallel pipeline.

[0017] In some embodiments, the pipeline system includes a third parallel pipeline, which is arranged in parallel with the first parallel pipeline and the second parallel pipeline. One end of the third parallel pipeline is connected to the outlet end of the rough sorting container, and the other end is connected to the intersection pipeline.

[0018] In some embodiments, a first branch is provided on the first parallel pipeline, and the first branch is provided in parallel with the at least one sorting container on the first parallel pipeline.

[0019] In some embodiments, a second branch is provided on the second parallel pipeline, and the second branch is provided in parallel with the at least one sorting container on the second parallel pipeline.

[0020] In some embodiments, the pipeline system further includes a third waste liquid bag and a fifth pipeline, and both ends of the fifth pipeline are connected to the third waste liquid bag and the junction pipeline respectively.

[0021] In some embodiments, the pipeline system further includes a first terminal pipeline, a second terminal pipeline, a first waste liquid bag, and a destination cell bag; wherein, the two ends of the first terminal pipeline are respectively connected to the intersection pipeline and the first waste liquid bag, and the two ends of the second terminal pipeline are respectively connected to the intersection pipeline and the destination cell bag.

[0022] In some embodiments, the pipeline system further includes a buffer bag and a buffer pipeline; wherein two ends of the buffer pipeline are respectively connected to the junction pipeline and the buffer bag.

[0023] In some embodiments, the pipeline system further includes a second waste liquid bag and a third pipeline; wherein both ends of the third pipeline are respectively connected to the coarse sorting container and the second waste liquid bag.

[0024] In some embodiments, the pipeline system further includes a sample bag, a fourth pipeline and a second bubble sensor, wherein two ends of the fourth pipeline are respectively connected to the sample bag and the coarse sorting container, and the second bubble sensor is disposed on the fourth pipeline.

[0025] In some embodiments, the pipeline system further includes a plurality of pumps; wherein each of the plurality of pumps is respectively arranged corresponding to the coarse sorting container and one of the sorting containers, and the pumping speed of the pump corresponding to the coarse sorting container is greater than the pumping speed of the pump corresponding to the sorting container.

[0026] An embodiment of this specification provides a cell sorting device, which includes the pipeline system described in this specification.

[0027] An embodiment of the present specification provides a control method for a cell sorting system, characterized in that the method includes: starting a cell sorting device and performing alternating sorting or simultaneous sorting using a plurality of sorting containers arranged in parallel.

[0028] In some embodiments, the method further comprises: performing a coarse sorting control operation or a filtration operation using a coarse sorting container before performing the alternating sorting or the simultaneous sorting.

[0029] In some embodiments, the coarse sorting control operation includes: starting a coarse sorting magnetic field assembly; controlling the cell sample to flow through a coarse sorting container, and collecting target cells flowing out of the coarse sorting container into a cell transfer bag.

[0030] In some embodiments, the cell sorting device includes a fourth pipeline and a second bubble sensor disposed on the fourth pipeline; the method further includes: stopping the coarse sorting control operation or the filtration operation based on the second bubble sensor detecting a trigger stop signal.

[0031] In some embodiments, the cell sorting device includes a second pipeline and a first bubble sensor disposed on the second pipeline; the method further includes: stopping the alternating sorting or the simultaneous sorting based on the first bubble sensor detecting a trigger stop signal. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The present application will be further described in the form of exemplary embodiments, which will be described in detail with reference to the accompanying drawings. These embodiments are not limiting, and in these embodiments, like numbers represent like structures, wherein:

[0033] FIG1 is a schematic structural diagram of a piping system for cell sorting according to some embodiments of this specification;

[0034] FIG2 is a schematic structural diagram of a piping system for cell sorting according to yet other embodiments of this specification;

[0035] FIG3 is a schematic structural diagram of a piping system for cell sorting according to yet other embodiments of this specification;

[0036] FIG4 is a schematic structural diagram of a sorting container according to some embodiments of this specification;

[0037] FIG5A is a schematic structural diagram of a piping system for cell sorting according to yet other embodiments of the present specification;

[0038] FIG5B is a schematic structural diagram of a pipeline system for cell sorting according to yet other embodiments of this specification;

[0039] FIG5C is a schematic structural diagram of a pipeline system for cell sorting according to yet other embodiments of this specification;

[0040] FIG6 is an exemplary flow chart of a method for controlling a cell sorting system according to some embodiments of the present specification. DETAILED DESCRIPTION

[0041] To more clearly illustrate the technical solutions of the embodiments of this application, the following is a brief introduction to the drawings required for describing the embodiments. Obviously, the drawings described below are merely examples or embodiments of this application. Those skilled in the art can apply this application to other similar scenarios based on these drawings without inventive effort. Unless otherwise apparent from the context or otherwise noted, the same reference numerals in the figures represent the same structure or operation.

[0042] It should be understood that the terms "system," "device," "unit," and / or "module" used herein are a method for distinguishing different components, elements, parts, portions, or assemblies at different levels. However, if other terms can achieve the same purpose, the terms may be replaced by other expressions.

[0043] As used in this application and the claims, unless the context clearly indicates otherwise, the words "a," "an," "an," and / or "the" are not intended to refer to the singular but may include the plural. Generally speaking, the terms "comprises" and "include" only indicate the inclusion of the steps and elements specifically identified, and these steps and elements do not constitute an exclusive list. A method or apparatus may also include other steps or elements.

[0044] The purpose of cell sorting is to obtain ideal target cells. High-quality cell sorting requires not only high purity and high yield of the target cells, but also reproducibility, as well as ensuring cell viability and normal cell function after sorting. In the actual cell sorting process, it is often necessary to process a large sample volume while also hoping for a shorter sorting time and less impact on cell activity. Existing immunomagnetic cell sorting equipment has a limited single sample sorting volume and a long sorting time. On the one hand, it is difficult to meet the sample volume requirements. On the other hand, it takes too long to run when sorting larger sample volumes, which has a greater impact on cell viability and normal cell function.

[0045] The embodiments of this specification provide a pipeline assembly for cell sorting. The pipeline assembly includes a magnetic field assembly and a plurality of sorting containers arranged in the magnetic field assembly, wherein the plurality of sorting containers are arranged in parallel. The cell sample is driven to flow in the pipeline system by a pump, so that the plurality of sorting containers arranged in parallel are alternately sorted or sorted simultaneously in the magnetic field module, thereby increasing the sample processing capacity, shortening the sorting time, reducing the reduction in cell activity and cell death caused by the long sorting time, breaking through the single sample sorting volume and sorting time limitations of existing equipment, and achieving high-quality cell sorting with large volume, high purity, high yield, reproducibility, and normal cell viability and cell function in a short time.

[0046] In some embodiments, the pipeline assembly may further include a coarse sorting container, which is arranged in series before the multiple sorting containers. In some embodiments, the coarse sorting container is a container used to perform a coarse sorting operation. The coarse sorting operation refers to the magnetic field component acting on the coarse sorting container, and the cell sample flows through the coarse sorting container at a relatively fast rate. In some embodiments, when the magnetic field component is not acting on the coarse sorting container, the coarse sorting container can be referred to as a filtration container or a pre-sorting container. At this time, the cell sample flows through the filtration container at a relatively fast rate to perform a filtration operation. Because the flow rate of the cell sample in the coarse sorting operation or filtration operation is relatively fast and rough, the obtained target cell solution or filtrate may still contain a large amount of impurities. Therefore, a fine sorting operation can also be performed after the coarse sorting operation or filtration operation. In some embodiments, the sorting container is a container used to perform a fine sorting operation. The sorting container can also be referred to as a fine sorting container. The fine sorting operation refers to allowing the sample solution to flow through the sorting container at a relatively slow rate. Because the fine sorting operation has a slower and more precise flow rate, it can more comprehensively remove non-target cells and other impurities, obtaining purer target cells. By combining the coarse sorting operation or filtration operation with the fine sorting operation, the cell sample is first subjected to the coarse sorting operation or filtration operation, and then the cell sample is subjected to the fine sorting operation, so that the purity of the sorted cells is higher and the cell sorting efficiency is improved.

[0047] The pipeline assembly provided in the embodiments of this specification will be described in detail below with reference to the accompanying drawings.

[0048] Figure 1 is a schematic diagram of the structure of a piping system for cell sorting according to some embodiments of this specification; Figure 2 is a schematic diagram of the structure of a piping system for cell sorting according to some other embodiments of this specification; Figure 3 is a schematic diagram of the structure of a piping system for cell sorting according to some other embodiments of this specification.

[0049] As shown in Figures 1-3, the piping system 100 may include a magnetic field assembly 110 and multiple sorting containers 120. The multiple sorting containers 120 may be disposed within the magnetic field assembly 110. In some embodiments, the multiple sorting containers 120 may be disposed in parallel. In some embodiments, as shown in Figures 1-2, N may represent the number of sorting containers 120, and N sorting containers 120 may be disposed in parallel. In some embodiments, N may be a positive integer greater than 1, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. In some embodiments, as shown in Figure 3, the multiple sorting containers 120 may be divided into multiple groups, such as Group A and Group B. In some embodiments, N1, N2, or N3 may represent the number of sorting containers 120 in one of the multiple groups, such as Group A having N1 sorting containers 120 and Group B having N2 sorting containers 120. In some embodiments, (N1+N2+N3) sorting containers 120 may be disposed in parallel. In some embodiments, N1, N2, or N3 may be a positive integer, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. In some embodiments, N1, N2, or N3 may be different, for example, N1 is 1, N2 is 2, and N3 is 4. In some embodiments, if N1, N2, and N3 are all 1, the piping system 100 shown in FIG3 is similar to the piping system 100 shown in FIG1 .

[0050] In some embodiments, the pipeline system 100 may further include a sample bag 130 and a parallel pipeline 140, and the parallel pipeline 140 may be arranged between the sample bag 130 and the plurality of sorting containers 120. In some embodiments, the number of sample bags 130 may be one or more. In some embodiments, when there are multiple sample bags 130, the multiple sample bags 130 contain the same type of cell samples, and the multiple sample bags 130 and the parallel pipeline 140 may be closed and connected. In some embodiments, as shown in Figures 1-2, the parallel pipeline 140 may include N parallel branch pipelines and sorting containers 120. In some embodiments, as shown in Figure 3, the parallel pipeline 140 may include (N1+N2+N3) parallel branch pipelines and sorting containers 120.

[0051] In some embodiments, as shown in Figures 1 and 3, the magnetic field assembly 110 may include a plurality of magnetic field modules. In some embodiments, as shown in Figure 1, the number of sorting containers 120 may be the same as the number of magnetic field modules, and each of the plurality of sorting containers 120 may be respectively arranged in a magnetic field module. In some embodiments, a plurality of sorting containers 120 may simultaneously perform a cell sorting operation in the magnetic field assembly 110. In some embodiments, performing a cell sorting operation simultaneously may be causing a cell sample to flow through a plurality of sorting containers 120 simultaneously to perform a cell sorting operation simultaneously. For example, as shown in Figure 1, a cell sample flows through N sorting containers 120 simultaneously to perform a cell sorting operation simultaneously. In some embodiments, at least two of the plurality of sorting containers 120 may simultaneously perform a cell sorting operation in the magnetic field assembly 110. In some embodiments, if more than two sorting containers 120 are included, the cell sample may be caused to flow through at least two of the sorting containers 120 simultaneously to perform a cell sorting operation simultaneously. For example, as shown in FIG1 , the cell sample flows through at least two of the N sorting containers 120 (where N is greater than 2) at the same time, so that at least two sorting containers 120 perform cell sorting operations at the same time. Compared with a single sorting container, when the cell sorting operation is performed by the parallel piping system shown in FIG1 , when N=3, three sorting containers 120 perform cell sorting operations at the same time. For the same amount of cell sample, the sorting time can be reduced by 67% through the parallel piping system shown in FIG1 , or within the same sorting time, the amount of sorted samples can be increased by 67% through the parallel piping system shown in FIG1 ; when N=4, four sorting containers 120 perform cell sorting operations at the same time. For the same amount of cell sample, the cell sorting operation can be performed by the parallel piping system shown in FIG1 . For the cell sorting operation, the sorting time of the parallel-arranged pipeline system shown in FIG1 can be reduced by 75%, or within the same sorting time, the sorting sample volume of the parallel-arranged pipeline system shown in FIG1 can be increased by 75%; when N=5, 5 sorting containers 120 perform the cell sorting operation at the same time, and for the same cell sample volume, the sorting time of the parallel-arranged pipeline system shown in FIG1 can be reduced by 80%, or within the same sorting time, the sorting sample volume of the parallel-arranged pipeline system shown in FIG1 can be increased by 80%.

[0052] In some embodiments, as shown in Figure 2, the magnetic field assembly 110 may include a magnetic field module, and multiple sorting containers 120 may share one magnetic field module. In some embodiments, one magnetic field module may be fixedly arranged, and multiple sorting containers 120 may be moved in sequence. After a sorting container 120 completes a cell sorting operation in the magnetic field module, the next sorting container 120 is moved into the magnetic field module to complete the next cell sorting operation, and so on and so forth to complete the cell sorting operation in multiple sorting containers 120. In some embodiments, multiple sorting containers 120 may be fixedly arranged, and one magnetic field module may be moved in sequence so that multiple sorting containers 120 are sequentially located in the magnetic field module. After a sorting container 120 completes a cell sorting operation in the magnetic field module, the magnetic field module is moved to the next sorting container 120 to complete the next cell sorting operation, and so on and so forth to complete the cell sorting operation in multiple sorting containers 120.

[0053] In some embodiments, as shown in FIG3 , the number of sorting containers 120 can be greater than the number of magnetic field modules, and at least two sorting containers 120 can share one magnetic field module. In some embodiments, as shown in FIG3 , group A or group B respectively includes at least two sorting containers 120 and one magnetic field module, and in group A or group B, at least two sorting containers 120 can share one magnetic field module. In some embodiments, taking group A as an example, the magnetic field module in group A can be fixed, and multiple sorting containers 120 can be moved in sequence. After a sorting container 120 completes the cell sorting operation in the magnetic field module, the next sorting container 120 is moved to the magnetic field module to complete the next cell sorting operation, and so on to complete the cell sorting operation in multiple sorting containers 120. In some embodiments, the multiple sorting containers 120 in group A can be fixedly arranged, and a magnetic field module is moved in sequence so that the multiple sorting containers 120 are located in the magnetic field module in sequence. After a sorting container 120 completes the cell sorting operation in the magnetic field module, the magnetic field module is moved to the next sorting container 120 to complete the next cell sorting operation, and so on to complete the cell sorting operation in the multiple sorting containers 120 in group A. In some embodiments, the multiple groups in Figure 3 (for example, Group A and Group B) can perform cell sorting operations at the same time. In some embodiments, the multiple groups in Figure 3 performing cell sorting operations simultaneously can be to make the cell samples flow through the sorting containers 120 in the magnetic field module in the multiple groups (for example, Group A and Group B) at the same time, so that the multiple groups perform cell sorting operations at the same time. In some embodiments, the multiple groups in Figure 3 (for example, Group A and Group B) can perform cell sorting operations in sequence. In some embodiments, the multiple groups in FIG3 sequentially perform cell sorting operations by allowing the cell sample to flow sequentially through the sorting container 120 in the magnetic field module of one of the groups (e.g., Group A or Group B), and so on and so forth to sequentially complete a cell sorting operation of the multiple groups. In some embodiments, after all the groups in FIG3 have sequentially performed the cell sorting operation, if the cell sorting operation has not been completed, the next sorting container 120 in each group can be moved to the magnetic field module or one of the magnetic field modules in each group can be moved so that the next sorting container 120 is located in the magnetic field module, and the cell sample is again sequentially passed through the sorting container 120 in the magnetic field module of one of the groups (e.g., Group A or Group B), and so on and so forth to sequentially complete the next cell sorting operation of the multiple groups. In some embodiments, when the multiple groups in FIG3 sequentially perform cell sorting operations, each group performs the cell sorting operation equivalent to the pipeline system in FIG2 performing the cell sorting operation.

[0054] Figure 4 is a schematic diagram of the structure of a sorting container according to some embodiments of the present disclosure. In some embodiments, the sorting container 120 can be used to adsorb cells labeled with magnetic particles to achieve the purpose of cell sorting.

[0055] As shown in FIG4 , the outer shell of the sorting container 120 comprises an upper shell 121 and a lower shell 122 , each of which is provided with a first open end 123 and a second open end 124 . The upper shell 121 and the lower shell 122 can form a cavity filled with a plurality of magnetic beads 125 , with gaps between the plurality of magnetic beads 125 allowing sample cells to pass through. In some embodiments, the magnetic beads 125 can comprise steel or iron balls, which become magnetic under the action of the magnetic field module. In some embodiments, the upper and lower ends of the cavity of the sorting container 120 are provided with a first sieve plate 126 and a second sieve plate 127 , respectively. The plurality of magnetic beads 125 are located within the cavity space confined by the first sieve plate 126 and the second sieve plate 127 to prevent the magnetic beads 125 from falling out of the sorting container 120 . In some embodiments, micropores are formed on the first sieve plate 126 and the second sieve plate 127 to facilitate the flow of fluid in the sorting container 120 .

[0056] Before the sample enters the sorting container 120, a magnetic field needs to be applied to the sorting container 120 to form a high magnetic gradient in the sorting container 120, thereby helping to adsorb the cells labeled with magnetic particles in the sample into the sorting container 120. The cells labeled with magnetic particles can be target cells or non-target cells. When the cells labeled with magnetic particles flow through the sorting container 120 from the first opening end 123, they can be adsorbed by the magnetic field with a high magnetic gradient and suspended between the gaps between the several magnetic beads 125, while the cells labeled with non-magnetic particles can pass smoothly between the gaps between the several magnetic beads 125 and flow out of the sorting container 120 through the second opening end 124. When the magnetic field is removed, the high magnetic gradient in the sorting container 120 disappears, which helps to release the cells labeled with magnetic particles adsorbed in the sorting container 120, thereby achieving the purpose of cell sorting.

[0057] Figure 5 is a structural schematic diagram of a piping system for cell sorting according to some other embodiments of the present specification; Figure 5B is a structural schematic diagram of a piping system for cell sorting according to some other embodiments of the present specification; Figure 5C is a structural schematic diagram of a piping system for cell sorting according to some other embodiments of the present specification.

[0058] In some embodiments, the cell sorting operation may include a coarse sorting operation and a fine sorting operation. In some embodiments, the coarse sorting operation may be performed before the fine sorting operation. In some embodiments, the sorting container corresponding to the fine sorting operation may include one sorting container. In some embodiments, the sorting container corresponding to the fine sorting operation may include multiple sorting containers arranged in parallel. By combining the coarse sorting operation and the fine sorting operation, the purity of the sorted cells can be increased, thereby improving the efficiency of cell sorting.

[0059] As shown in Figures 5A-5C, the pipeline system 100 may further include a coarse sorting container 150 (for example, coarse sorting container I), and the coarse sorting container 150 may be arranged in series before a plurality of sorting containers 120 (for example, sorting containers II, III, IV, V). In some embodiments, the number of coarse sorting containers 150 may be one or more. In some embodiments, a plurality of coarse sorting containers 150 may be arranged in parallel or in series. In some embodiments, the pipeline system 100 may further include a coarse sorting magnetic field assembly, and the coarse sorting container 150 may be arranged in the coarse sorting magnetic field assembly. The coarse sorting magnetic field assembly is similar to the magnetic field assembly 110 in Figures 1-3. For more information about the coarse sorting magnetic field assembly, please refer to the description of the magnetic field assembly 110 in Figures 1-3, which will not be repeated here.

[0060] In some embodiments, as shown in Figures 5A-5B, the pipeline system 100 may further include a cell transfer bag 161, a first pipeline 171, and a second pipeline 172. In some embodiments, the cell transfer bag 161 can be used to store cell samples processed by the rough sorting operation. In some embodiments, the two ends of the first pipeline 171 can be connected to the outlet end of the rough sorting container 150 and the first interface of the cell transfer bag 161, respectively, for transferring the cell sample processed by the rough sorting operation in the rough sorting container 150 to the cell transfer bag 161. In some embodiments, the first interface of the cell transfer bag 161 can be an inlet for the cell sample processed by the rough sorting operation. In some embodiments, a valve (e.g., valve 5 shown in Figures 5A-5B) can be provided on the first pipeline 171, and by opening or closing the valve, the fluid is allowed to flow through the first pipeline 171 or the fluid is prevented from flowing through the first pipeline 171. In some embodiments, the second conduit 172 can be disposed between the second interface of the cell transfer bag 161 and the inlet ends of the plurality of sorting containers 120, for transporting the cell sample stored in the cell transfer bag 161 to the plurality of sorting containers 120 for fine sorting. In some embodiments, the second interface of the cell transfer bag 161 can be an outlet for the cell sample stored in the cell transfer bag 161. In some embodiments, the inlet ends of the plurality of sorting containers 120 can be an inlet for the cell sample flowing into the sorting containers 120. In some embodiments, a valve (e.g., valve 8 shown in Figures 5A-5B) can be disposed on the second conduit 172, and by opening or closing the valve, the fluid is allowed to flow through the second conduit 172 or prevented from flowing through the second conduit 172.

[0061] In some embodiments, as shown in Figures 5A-5B, the piping system 100 may further include a first bubble sensor 181, which can be used to detect the presence of fluid in the piping. In some embodiments, the first bubble sensor 181 may be disposed on the second piping 172. In some embodiments, when the first bubble sensor 181 detects that there is no fluid in the second piping 172 (i.e., the liquid in the cell transfer bag 161 has been exhausted), it may send a detection signal to the control device to stop the sub-sorting operation.

[0062] In some embodiments, as shown in FIG5A , the parallel pipelines of the pipeline system 100 may include a first parallel pipeline 173-1, a second parallel pipeline 173-2, and a junction pipeline 174. In some embodiments, the first parallel pipeline 173-1 and the second parallel pipeline 173-2 may be arranged in parallel. In some embodiments, one end of the first parallel pipeline 173-1 and the second parallel pipeline 173-2 may be connected to the outlet of the rough sorting container 150. In some embodiments, the other ends of the first parallel pipeline 173-1 and the second parallel pipeline 173-2 may be connected to the junction pipeline 174 and located before the junction pipeline 174. In some embodiments, a valve (e.g., valve 9 shown in FIG5A-5B ) may be provided on the first parallel pipeline 173-1. By opening or closing the valve, the fluid is allowed to flow through the first parallel pipeline 173-1 or prevented from flowing through the first parallel pipeline 173-1. In some embodiments, a valve (e.g., valve 10 as shown in Figures 5A-5B) may be provided on the second parallel line 173-2. By opening or closing the valve, the fluid is allowed to flow through the second parallel line 173-2 or prevented from flowing through the second parallel line 173-2. By simultaneously providing the first parallel line 173-1 and the second parallel line 173-2, the fluid flowing through the second line 172 may be dispersed into at least three lines. In some embodiments, a sorting container 120 may be provided on the first parallel line 173-1. In some embodiments, at least two sorting containers 120 may be provided on the first parallel line 173-1, and at least two sorting containers 120 may be provided in parallel. In some embodiments, a sorting container 120 may be provided on each of the second parallel lines 173-2. In some embodiments, at least two sorting containers 120 may be provided on the second parallel line 173-2, and at least two sorting containers 120 may be provided in parallel. By arranging at least one sorting container on the first parallel pipeline and / or the second parallel pipeline, multiple fluid paths for cell subdivision sorting operations can be formed. The multiple fluid paths all pass through the first parallel pipeline and / or the second parallel pipeline and the sorting container to realize various steps in the cell subdivision sorting operation.

[0063] In some embodiments, as shown in Figures 5B-5C, a first branch 173-4 may be provided on the first parallel line 173-1. In some embodiments, the first branch 173-4 may be provided in parallel with one or more sorting containers 120 on the first parallel line 173-1. In some embodiments, a valve (e.g., valve 17 as shown in Figures 5B-5C) may be provided on the first branch 173-4, and by opening or closing the valve, the fluid is allowed to flow through the first branch or prevented from flowing through the first branch. In some embodiments, as shown in Figures 5B-5C, a second branch 173-5 may be provided on the second parallel line 173-2. In some embodiments, the second branch 173-5 may be provided in parallel with one or more sorting containers 120 on the second parallel line 173-2. In some embodiments, a valve (e.g., valve 18 as shown in Figures 5B-5C) may be provided on the second branch 173-5, and by opening or closing the valve, the fluid is allowed to flow through the second branch or prevented from flowing through the second branch. By setting up the first branch and the second branch, the sorting containers on the first parallel pipeline and the second parallel pipeline can be flushed at the same time. For example, the sorting container II on the first parallel pipeline and the sorting container IV on the second parallel pipeline can be flushed at the same time, and then the sorting container III on the first parallel pipeline and the sorting container V on the second parallel pipeline can be flushed at the same time without flushing the sorting containers one by one. The flushing efficiency can be improved (for example, the flushing efficiency of the pipeline system shown in Figures 5B-5C is twice that of the pipeline system shown in Figure 5A), and the total time of the sorting and flushing operations can be shortened.

[0064] In some embodiments, as shown in FIG5A , the parallel pipelines of the pipeline system 100 may include a third parallel pipeline 173-3. In some embodiments, the third parallel pipeline 173-3 may be arranged in parallel with the first parallel pipeline 173-1 and the second parallel pipeline 173-2. In some embodiments, one end of the third parallel pipeline 173-3 may be connected to the outlet of the coarse sorting container 150, and the other end may be connected to the junction pipeline 174. In some embodiments, a valve (e.g., valve 11 shown in FIG5A ) may be provided on the third parallel pipeline 173-3. By opening or closing the valve, the fluid is allowed to flow through the third parallel pipeline 173-3 or prevented from flowing through the third parallel pipeline 173-3. By providing a third parallel pipeline, the second pipeline can be directly connected to the junction pipeline, without performing a fine sorting operation on the sample cells.

[0065] In some embodiments, as shown in Figures 5A-5C , a junction line 174 can be disposed after the first parallel line 173-1 and the second parallel line 173-2. In some embodiments, a valve (e.g., valve 13 as shown in Figures 5A-5C ) can be disposed on the junction line 174 to allow or prevent fluid from flowing through the junction line 174 by opening or closing the valve. The junction line 174 can combine the fluids flowing through the first parallel line 173-1, the second parallel line 173-2, and the third parallel line 173-3 into a single line.

[0066] In some embodiments, as shown in Figures 5A-5B, the tubing system 100 may further include a first terminal tubing 175, a second terminal tubing 176, a first waste liquid bag 191, and a destination cell bag 162. In some embodiments, the two ends of the first terminal tubing 175 may be connected to the junction tubing 174 and the first waste liquid bag 191, respectively. In some embodiments, the first waste liquid bag 191 may be an independent cell bag or a waste liquid bag. In some embodiments, a valve (e.g., valve 14 as shown in Figures 5A-5C) may be provided on the first terminal tubing 175 to allow fluid to flow through the first terminal tubing 175 or to prevent fluid from flowing through the first terminal tubing 175 by opening or closing the valve. Through the first terminal tubing 175, the fluid flowing through the junction tubing 174 may be collected in the first waste liquid bag 191. In some embodiments, the two ends of the second terminal tubing 176 may be connected to the junction tubing 174 and the destination cell bag 162, respectively. In some embodiments, a valve (e.g., valve 15 as shown in Figures 5A-5C) can be provided on the second terminal tube 176, and by opening or closing the valve, the fluid is allowed to flow through the second terminal tube 176 or prevented from flowing through the second terminal tube 176. The fluid flowing through the junction tube 174 can be collected in the target cell bag 162 through the second terminal tube 176.

[0067] In some embodiments, as shown in Figures 5A-5C, the piping system 100 may further include a buffer bag 193 and a buffer line 177. In some embodiments, the two ends of the buffer line 177 may be connected to the junction line 174 and the buffer bag 193, respectively. In some embodiments, a valve (e.g., valve 12 as shown in Figures 5A-5C) may be provided on the buffer line 177. By opening or closing the valve, the fluid is allowed to flow through the buffer line 177 or prevented from flowing through the buffer line 177. Through the buffer bag, the buffer line, and the piping related to the cell transfer bag, the buffer in the buffer bag can be filled into the cell transfer bag to wash the residual cells in the cell transfer bag. In some embodiments, the composition of the buffer may be different in different application scenarios. The buffer may include at least PBS (Phosphate Buffered Saline) buffer and ethylenediaminetetraacetic acid (EDTA).

[0068] In some embodiments, as shown in Figures 5A-5C , the piping system 100 may further include a first intermediate piping 177-1. In some embodiments, the first intermediate piping 177-1 may be connected at both ends to the buffer bag 193 and the fourth piping 179, respectively. In some embodiments, a valve (e.g., valve 1 as shown in Figures 5A-5C ) may be provided on the first intermediate piping 177-1. Opening or closing the valve allows or blocks fluid flow through the first intermediate piping. The provision of the first intermediate piping allows the buffer bag to be directly connected to the fourth piping, facilitating cleaning of the coarse sorting container 150.

[0069] In some embodiments, as shown in Figures 5A-5C, the piping system 100 may further include a second intermediate pipeline 177-2. In some embodiments, the ends of the second intermediate pipeline 177-2 may be connected to the buffer bag 193 and the first parallel pipeline 173-1, the second parallel pipeline 173-2, or the third parallel pipeline 173-3, respectively. In some embodiments, a valve (e.g., valve 6 as shown in Figures 5A-5C) may be provided on the second intermediate pipeline 177-2. By opening or closing the valve, the fluid is allowed to flow through the second intermediate pipeline or prevented from flowing through the second intermediate pipeline. By providing the second intermediate pipeline, the buffer bag can be directly connected to the first parallel pipeline, the second parallel pipeline, or the third parallel pipeline, facilitating the cleaning of the sorting container 120.

[0070] In some embodiments, as shown in Figures 5A-5C, the piping system 100 may further include a third intermediate pipeline 177-3. In some embodiments, the ends of the third intermediate pipeline 177-3 may be connected to the coarse sorting container 150 and the first parallel pipeline 173-1, the second parallel pipeline 173-2, or the third parallel pipeline 173-3, respectively. In some embodiments, a valve (e.g., valve 7 as shown in Figures 5A-5C) may be provided on the third intermediate pipeline 177-3. By opening or closing the valve, the fluid is allowed to flow through the third intermediate pipeline or prevented from flowing through the third intermediate pipeline. By providing the third intermediate pipeline, the coarse sorting container can be directly connected to the first parallel pipeline, the second parallel pipeline, or the third parallel pipeline, allowing the fine sorting operation to be directly performed after the coarse sorting operation.

[0071] In some embodiments, as shown in Figures 5A-5B, the piping system 100 may further include a fourth intermediate pipeline 177-4. In some embodiments, the ends of the fourth intermediate pipeline 177-4 may be connected to the second intermediate pipeline 177-2 or the third intermediate pipeline 177-3 and the second pipeline 172, respectively. In some embodiments, a valve (e.g., valve 16 shown in Figure 5B) may be provided on the fourth intermediate pipeline 177-4. By opening or closing the valve, the flow of fluid through the fourth intermediate pipeline is allowed or prevented.

[0072] By arranging a valve 16 on the fourth intermediate pipeline, during the fine sorting operation, if the coarse sorting operation has been completed, the valve 16 can be closed to flush the coarse sorting container and related pipelines performing the coarse sorting operation during the fine sorting operation without hindering the continuation of the fine sorting operation, thereby improving the sorting efficiency and shortening the total time of the sorting operation.

[0073] In some embodiments, as shown in Figures 5A-5B, the pipeline system 100 may further include a second waste liquid bag 192 and a third pipeline 178. In some embodiments, the two ends of the third pipeline 178 may be connected to the rough sorting container 150 and the second waste liquid bag 192, respectively. In some embodiments, the second waste liquid bag 192 may be an independent cell bag or a waste liquid bag. In some embodiments, a valve (for example, valve 4 as shown in Figures 5A-5B) may be provided on the third pipeline 178, and by opening or closing the valve, the fluid is allowed to flow through the third pipeline 178 or the fluid is prevented from flowing through the third pipeline 178. Through the third pipeline, the fluid after the rough sorting operation in the rough sorting container can be collected in the second waste liquid bag.

[0074] In some embodiments, as shown in Figures 5A-5C , the piping system 100 may further include a sample bag 130 and a fourth piping 179. In some embodiments, the ends of the fourth piping 179 may be connected to the sample bag 130 and the coarse sorting container 150, respectively. In some embodiments, a valve (e.g., valve 2 shown in Figures 5A-5C ) may be provided on the fourth piping 179. By opening or closing the valve, the valve allows or blocks fluid flow through the fourth piping 179. Through the fourth piping, sample cells in the sample bag can be flowed into the coarse sorting container for coarse sorting. In some embodiments, the piping system 100 may further include a second bubble sensor 182. In some embodiments, the second bubble sensor 182 may be provided on the fourth piping 179. In some embodiments, when the second bubble sensor 182 detects that there is no fluid in the fourth piping 179 (i.e., the liquid in the sample bag 130 has flowed out), it may send a detection signal to the control device to stop the coarse sorting operation.

[0075] In some embodiments, the piping system 100 may further include a fifth intermediate pipeline 177-5. In some embodiments, the ends of the fifth intermediate pipeline 177-5 may be connected to the fourth pipeline 179 and the third pipeline 178, respectively. In some embodiments, a valve (e.g., valve 3 as shown in Figures 5A-5B) may be provided on the fifth intermediate pipeline 177-5. By opening or closing the valve, the valve allows or blocks fluid flow through the fifth intermediate pipeline 177-5. By providing the fifth intermediate pipeline 177-5, the fourth pipeline and the third pipeline can be directly connected, eliminating the need to clean the rough sorting container 150.

[0076] In some embodiments, as shown in FIG. 5C , a valve (eg, valve 3 ) may be provided on the third pipeline 178 , and both ends of the third pipeline 178 may be connected to the fourth pipeline 179 and the second waste liquid bag 192 , respectively.

[0077] In some embodiments, as shown in FIG5C , the tubing system 100 may include a fifth tubing 170 and a third waste liquid bag 194. In some embodiments, the ends of the fifth tubing 170 may be connected to the junction tubing 174 and the third waste liquid bag 194, respectively. By providing the fifth tubing and the third waste liquid bag, the solution that flows through can be collected into the third waste liquid bag 194 during the fine separation operation.

[0078] In some embodiments, the piping system 100 may further include multiple pumps (not shown). In some embodiments, the multiple pumps may be non-contact pumps, such as peristaltic pumps. In some embodiments, each of the multiple roughing containers 150 may be provided with a pump. In some embodiments, each of the one or more roughing containers 150 may be provided with a pump. The pump corresponding to the roughing container may be referred to as a roughing pump. In some embodiments, the number of roughing containers 150 may be greater than or equal to the number of roughing pumps. In some embodiments, each of the multiple sorting containers 120 may be provided with a pump. The pump corresponding to each sorting container may be referred to as a fine sorting pump. In some embodiments, the number of the multiple sorting containers 120 may be greater than or equal to the number of fine sorting pumps. In some embodiments, the number of pumps may be equal to the sum of the number of roughing containers 150 and the multiple sorting containers 120. By setting the number of pumps equal to the sum of the number of roughing containers and the number of sorting containers, each roughing container and each sorting container may be provided with a pump. The pumps drive the flow of liquid in the pipeline, thereby increasing the liquid flow rate and improving sorting efficiency.

[0079] In some embodiments, the pumping speed of the pump corresponding to the coarse sorting container 150 can be greater than the pumping speed of the pump corresponding to the sorting container 120. By setting the pumping speed of the pump corresponding to the coarse sorting container to be greater, the flow rate and flow rate of the driven liquid can meet the required flow rate and flow rate of the liquid in multiple sorting containers arranged in parallel.

[0080] In some embodiments, each step in the cell sorting process is achieved by fluids (e.g., sample fluid and / or buffer fluid) flowing along corresponding fluid paths. Below, some key cell sorting processes are described in detail with reference to the various piping systems 100 shown in Figures 5A-5C.

[0081] In some embodiments, a cell sorting process based on the tubing system 100 shown in FIG. 5A may include the following steps:

[0082] Step 1: Preparation before sorting. The main operating procedures include: after starting the sorting equipment, filling or priming the piping system 100 with buffer to remove air from the piping; and cleaning the coarse sorting container 150 and the sorting container 120. During this process, the magnetic field assembly 110 does not act on the coarse sorting container 150 and the sorting container 120.

[0083] Step 2: Coarse sorting operation. The main operation process includes: applying the magnetic field assembly 110 to the coarse sorting container 150, opening valve 2 on the fourth pipeline 179 and valve 5 on the first pipeline 171, causing the liquid in the sample bag 130 to flow through the coarse sorting container 150. The peristaltic pump corresponding to the coarse sorting container 150 operates at a relatively fast and uniform speed. At this time, non-target cells labeled with magnetic particles are adsorbed by the coarse sorting container 150, and target cells that flow through the coarse sorting container 150 are collected in the cell transfer bag 161. When the second bubble sensor 182 detects that the liquid in the sample bag 130 has flowed out, it sends a detection signal to the control component, which stops the coarse sorting operation process.

[0084] Step 3, fine sorting operation. In some embodiments, the fine sorting operation can be performed in parallel after a period of time (for example, 1 minute, 3 minutes, or 5 minutes) after the coarse sorting operation begins. The main operation process of the fine sorting operation includes: allowing the magnetic field component 110 to act on the sorting container 120 (for example, sorting container II, III, IV, V), respectively opening the valve 8 on the second pipeline 172, the valve 9 on the first parallel pipeline 173-1, the valve 10 on the second parallel pipeline 173-2, the valve 13 on the intersection pipeline 174, and the valve 15 on the second terminal pipeline 176, and performing further fine sorting operations on the liquid in the cell transfer bag 161. At this time, the peristaltic pumps corresponding to the sorting containers 120 (e.g., sorting containers II, III, IV, V) operate at a slow and uniform speed, and the liquid in the cell transfer bag 161 is evenly pumped into the sorting containers 120 (e.g., sorting containers II, III, IV, V). During this process, the remaining labeled non-target cells in the liquid are adsorbed by the sorting containers II, III, IV, V, and the target cells that flow through the sorting containers 120 are collected in the target cell bag 162. When the first bubble sensor 181 detects that the liquid in the cell transfer bag 161 has flowed out, it sends a detection signal to the control component, which stops the subdivision sorting operation process.

[0085] Step 4: Backflushing the cell transfer bag. The main operational process involves applying magnetic field assembly 110 to each separation container 120, opening valve 8 on second line 172, valve 9 on first parallel line 173-1 or valve 10 on second parallel line 173-2, valve 12 on buffer line 177, and valve 13 on junction line 174, respectively. This allows at least a portion of the buffer in buffer bag 193 to be filled into cell transfer bag 161 as backflushing liquid, thereby washing away any remaining target cells in cell transfer bag 161. Furthermore, based on the previous process, valve 15 on second terminal line 176 is opened to perform a fine sorting operation on the backflushing liquid containing the target cells in cell transfer bag 161.

[0086] Step 5: Flushing the sorting containers and pipelines. The main operating procedures include: applying the magnetic field assembly 110 to the coarse sorting container 150 and each sorting container 120, opening valve 1 on the pipeline between the buffer bag 193 and the sample bag and valve 4 on the third pipeline 178, respectively, flushing the coarse sorting container 150 and the fourth pipeline 179 in which it resides, and flushing any remaining cells in the fourth pipeline 179 into the second waste liquid bag 192. Then, opening valve 6 on the pipeline between the buffer bag 193 and the second pipeline 172, valve 9 on the first parallel pipeline 173-1, valve 13 on the junction pipeline 174, and valve 14 on the first terminal pipeline 175, respectively, and operating the peristaltic pumps corresponding to sorting containers II and III in sequence to flush sorting containers II, III, and the first parallel pipeline 173-1 in which they reside, and flushing any remaining cells in the pipelines into the first waste liquid bag 191. Accordingly, valve 6 on the pipeline between the buffer bag 193 and the second pipeline 172, valve 10 on the second parallel pipeline 173-2, valve 13 on the intersection pipeline 174 and valve 14 on the first terminal pipeline 175 are opened respectively, and the peristaltic pumps corresponding to the sorting containers IV and V are operated in sequence to flush the sorting containers IV, V and the second parallel pipeline 173-2 in which they are located, and the cells remaining in the pipeline are flushed into the first waste liquid bag 191.

[0087] Step 6: Release the non-target cells adsorbed in the sorting container and flush the pipeline. The main operating process includes: removing the magnetic field assembly 110 from all sorting containers (coarse sorting container 150 and sorting container 120), opening valve 1 on the pipeline between the buffer bag 193 and the sample bag, valve 4 on the third pipeline 178, and the peristaltic pump corresponding to the coarse sorting container 150, flushing the coarse sorting container 150 and the fourth pipeline where it is located, and flushing the previously adsorbed non-target cells into the second waste liquid bag 192. Then, opening valve 6 on the pipeline between the buffer bag 193 and the second pipeline 172, valve 9 on the first parallel pipeline 173-1, valve 13 on the junction pipeline 174, valve 14 on the first terminal pipeline 175, and the peristaltic pump corresponding to sorting container II, and flushing sorting container II and the corresponding pipeline (first parallel pipeline 173-1), and flushing the previously adsorbed non-target cells into the first waste liquid bag 191. Accordingly, valve 6 on the pipeline between the buffer bag 193 and the second pipeline 172, valve 9 on the first parallel pipeline 173-1, valve 13 on the junction pipeline 174, valve 14 on the first terminal pipeline 175, and the peristaltic pump corresponding to the sorting container III are opened, respectively, to flush the sorting container III and the corresponding pipeline (first parallel pipeline 173-1), and the previously adsorbed non-target cells are flushed into the first waste liquid bag 191. Then, valve 6 on the pipeline between the buffer bag 193 and the second pipeline 172, valve 10 on the second parallel pipeline 173-2, valve 13 on the junction pipeline 174, valve 14 on the first terminal pipeline 175, and the peristaltic pump corresponding to the sorting container IV are opened, and the sorting container IV and the corresponding pipeline (second parallel pipeline 173-2) are flushed, and the previously adsorbed non-target cells are flushed into the first waste liquid bag 191. Then, open valve 6 on the pipeline between the buffer bag 193 and the second pipeline 172, valve 10 on the second parallel pipeline 173-2, valve 13 on the intersection pipeline 174, valve 14 on the first terminal pipeline 175 and the peristaltic pump corresponding to the sorting container V respectively, flush the sorting container V and the corresponding pipeline (second parallel pipeline 173-2), and flush the previously adsorbed non-target cells into the first waste liquid bag 191.

[0088] In some embodiments, a cell sorting process based on the tubing system 100 shown in FIG. 5B may include the following steps:

[0089] Step 1: Preparation before sorting: For the description of step 1 in the cell sorting process shown in FIG5B , please refer to the description of step 1 in the cell sorting process shown in FIG5A , and will not be repeated here.

[0090] Step 2: Rough sorting operation: For the description of step 2 in the cell sorting process shown in FIG5B , please refer to the description of step 2 in the cell sorting process shown in FIG5A , and will not be repeated here.

[0091] Step 3: Sorting operation: For the description of step 3 in the cell sorting process shown in FIG5B , please refer to the description of step 3 in the cell sorting process shown in FIG5A , and will not be repeated here.

[0092] Step 4: Backflushing the cell transfer bag. For the description of step 4 in the cell sorting process shown in FIG5B , please refer to the description of step 4 in the cell sorting process shown in FIG5A , which will not be repeated here.

[0093] In some embodiments, valve 16 may be closed while steps three and four are performed simultaneously.

[0094] Step 5: Flushing the sorting containers and pipelines. The main operating procedures include: simultaneously opening valves 9, 10, 17, and 18, and simultaneously operating the pumps corresponding to sorting containers II and IV, flushing the corresponding sorting containers II and IV in the two independent circulation pipelines of the first parallel pipeline 173-1 and the second parallel pipeline 173-2. After the flushing is completed, the pumps are stopped; then, the pumps corresponding to sorting containers III and V are simultaneously operated, flushing the corresponding sorting containers III and V in the two independent circulation pipelines of the first parallel pipeline 173-1 and the second parallel pipeline 173-2. In some embodiments, the order in which the pumps corresponding to the sorting containers are operated simultaneously can be any combination. For example, the pumps corresponding to sorting containers II and V can be operated simultaneously first, followed by the pumps corresponding to sorting containers III and IV. For another example, the pumps corresponding to sorting containers III and V can be operated simultaneously first, followed by the pumps corresponding to sorting containers II and IV.

[0095] Step 6: Release the non-target cells adsorbed in the sorting container and flush the pipeline. The description of step 6 in the cell sorting process shown in Figure 5B can be found in the description of step 6 in the cell sorting process shown in Figure 5A, and will not be repeated here.

[0096] In some embodiments, a cell sorting process based on the tubing system 100 shown in FIG. 5C may include the following steps:

[0097] Step 1: Preparation before sorting. The main operation process includes: after starting the sorting equipment, filling or perfusing the pipeline system 100 with buffer to remove air in the pipeline and clean the coarse sorting container 150 and the sorting container 120; during this process, the magnetic field component 110 does not act on the coarse sorting container 150 and the sorting container 120. It should be noted that in the pipeline system 100 shown in Figure 5C, since the magnetic field component 110 does not act on the coarse sorting container 150 during the entire process, the coarse sorting container 150 does not perform the coarse sorting operation process and only serves as a filtering container or pre-sorting container. The coarse sorting container 150 can also be called a filtering container or a pre-sorting container.

[0098] Step 2, cell sorting operation, includes pre-sorting operation and fine sorting operation. The main operation process includes: allowing the magnetic field assembly 110 to act on the sorting container 120, without acting on the coarse sorting container 150 (no coarse sorting operation, only filtering operation, serving as the container for pre-sorting operation), opening valves 2, 7, 9, 10, 13, and 19, filtering the sample solution in the sample bag 130 through the coarse sorting container 150, and then flowing it into the sorting container 120 for sorting. At this time, the peristaltic pump corresponding to the sorting container 120 runs at a relatively slow and uniform speed, and the liquid in the sample bag 130 is evenly pumped through the sorting container 120. During this process, the labeled target cells in the sample solution are adsorbed by the sorting container 120, and the solution that flows through is collected in the third waste liquid bag 194; when the second bubble sensor 182 detects that the liquid in the sample bag 130 has flowed out, the cell sorting process is stopped.

[0099] Step 3: Backflushing the sample bag. The main operating process includes: applying magnetic field assembly 110 to each separation container 120, opening valves 2, 7, 9, 10, 12, and 13, and using a peristaltic pump to pump a portion of buffer into sample bag 130 to clean residual cells from sample bag 130 and related pipelines. Then, valves 2, 7, 9, 10, 13, and 19 are opened again, and step 2 is repeated to perform another separation operation on the backflushed liquid containing the target cells that has been backflushed into sample bag 130.

[0100] In some embodiments, the operation of step 3 may be repeated multiple times, and each cycle is stopped when the second bubble sensor 182 detects that the liquid in the sample bag 130 has flowed out.

[0101] Step 4: Rinse the sorting container and related pipelines. The main operation process includes: allowing the magnetic field assembly 110 to act on each sorting container 120, opening valve 1 and valve 3, flushing the pipeline, and the flushing liquid flows into the second waste liquid bag 192. Then, open valve 1, valve 7, valve 9, valve 10, valve 17, valve 18, valve 13, valve 14, flush the coarse sorting container 150 and related pipelines, and the flushing liquid flows into the first waste liquid bag 191. Then, open valve 6, valve 9, valve 13, valve 15, run the peristaltic pump corresponding to the sorting container 120 (for example, sorting container II) on the first parallel pipeline 173-1, flush the sorting container II on the first parallel pipeline 173-1 and its pipeline, and the flushing liquid flows into the first waste liquid bag 191. Then, valves 6, 9, 13, and 14 are opened, and the peristaltic pump corresponding to the sorting container 120 (e.g., sorting container III) on the first parallel pipeline 173-1 is operated to flush sorting container III and the pipeline in which it is located, and the flushing liquid flows into the first waste liquid bag 191. Then, valves 6, 10, 13, and 14 are opened, and the peristaltic pump corresponding to the sorting container 120 (e.g., sorting container IV) on the second parallel pipeline 173-2 is operated to flush sorting container IV and the pipeline in which it is located, and the flushing liquid flows into the first waste liquid bag 191. Then, valves 6, 10, 13, and 14 are opened, and the peristaltic pump corresponding to the sorting container 120 (e.g., sorting container V) on the second parallel pipeline 173-2 is operated to flush sorting container V and the pipeline in which it is located, and the flushing liquid flows into the first waste liquid bag 191.

[0102] In some embodiments, the order of flushing the sorting containers II, III, IV, and V can be arbitrarily changed.

[0103] Step 5: Release the target cells. The main operation process includes: removing the magnetic field assembly 110 corresponding to the sorting container 120 (for example, sorting containers II, III, IV, V); then opening valve 17 and valve 18, and simultaneously running the peristaltic pump corresponding to any one of the sorting containers 120 (for example, sorting containers II and III) on the first parallel pipeline 173-1, and the peristaltic pump corresponding to any one of the sorting containers 120 (for example, sorting containers IV and V) on the second parallel pipeline 173-2, to utilize The solution within this section of the pipeline is repeatedly flushed to remove the adsorbed target cells within the sorting container 120. The peristaltic pump corresponding to another sorting container (e.g., sorting containers II or III) in the first parallel pipeline 173-1 and the peristaltic pump corresponding to another sorting container (e.g., sorting containers IV or V) in the second parallel pipeline 173-2 are then operated to repeatedly flush the adsorbed target cells within the sorting container 120 using the solution within this section of the pipeline. Next, valves 6, 9, 10, 17, 18, 13, and 15 are opened to flush the target cells within this section of the pipeline into the target cell bag 162. Next, valves 6, 9, 13, and 15 are opened to operate the peristaltic pump corresponding to sorting container II, flushing sorting container II and the pipeline in which it resides, with the flushing solution flowing into the target cell bag 162. Then, open valves 6, 9, 13, and 15, and operate the peristaltic pump corresponding to sorting container III to flush sorting container III and the surrounding pipeline, allowing the flushing solution to flow into the target cell bag 162. Then, open valves 6, 10, 13, and 15, and operate the peristaltic pump corresponding to sorting container IV to flush sorting container IV and the surrounding pipeline, allowing the flushing solution to flow into the target cell bag 162. Then, open valves 6, 10, 13, and 15, and operate the peristaltic pump corresponding to sorting container V to flush sorting container V and the surrounding pipeline, allowing the flushing solution to flow into the target cell bag 162.

[0104] In some embodiments, the order of flushing the sorting containers II, III, IV, and V can be arbitrarily changed.

[0105] Some embodiments of this specification also provide a cell sorting device, which includes the piping system described in this specification. More technical details about the piping system can be found in the relevant descriptions of Figures 1 to 5 and will not be repeated here.

[0106] In some embodiments, the piping system shown in Figures 1 to 5 can be manufactured independently of the cell sorting apparatus. During use, the piping system can be installed on the cell sorting apparatus to perform the cell sorting process and can be removed after the cell sorting is completed. In some embodiments, the piping system shown in Figures 1 to 5 can be manufactured and used integrally with the cell sorting apparatus.

[0107] In some embodiments, the cell sorting device may further include a control component, which is used to control the pipeline system shown in Figures 1 to 5 to perform a cell sorting process.

[0108] FIG6 is an exemplary flow chart of a method for controlling a cell sorting system according to some embodiments of the present specification.

[0109] In some embodiments, process 600 may be performed by a processing device (or control component). For example, process 600 may be stored in a storage device in the form of a program or instructions, and when the processing device executes the program or instructions, process 600 may be implemented. In some embodiments, process 600 may utilize one or more additional operations not described below, and / or may not be completed by one or more operations discussed below. In addition, the order of the operations shown in FIG6 is not limiting.

[0110] Step 610: Start the cell sorting device.

[0111] In some embodiments, the cell sorting device can be started by operating a control module (eg, a switch button, a start control on an interface).

[0112] In some embodiments, after the sorting device is started, preparatory operations before sorting can be performed. More details about the preparatory operations before sorting can be found in the above description (e.g., Figures 5A-5B and step 1), which will not be repeated here.

[0113] Step 620: Perform a coarse sorting control operation or a filtering operation.

[0114] In some embodiments, a coarse sorting control operation or a filtering operation may be performed before performing a fine sorting operation of alternating sorting or simultaneous sorting.

[0115] In some embodiments, a coarse sorting container can be used to perform a coarse sorting control operation. In some embodiments, the control component can control the coarse sorting magnetic field component, the coarse sorting container, and open the valve on the corresponding pipeline to perform a coarse sorting control operation using the coarse sorting container. Specifically, the control component can control the start of the coarse sorting magnetic field component, so that the coarse sorting magnetic field component acts on the coarse sorting container, and then control the cell sample to flow through the coarse sorting container, and collect the target cells flowing out of the coarse sorting container in a cell transfer bag. In some embodiments, the cell sorting device may include a fourth pipeline, and a second bubble sensor (e.g., a second bubble sensor 182) provided on the fourth pipeline. In some embodiments, the control component can stop executing the coarse sorting control operation based on the second bubble sensor detecting a trigger stop signal. For more information about the coarse sorting control operation, please refer to the above description (e.g., Figures 5A-5B and step 2), which will not be repeated here.

[0116] In some embodiments, a filtering operation can be performed using a filtering container. In some embodiments, the control component can control the filtering container and open the valve on the corresponding pipeline to perform a filtering operation using the filtering container. Specifically, the control component can control the cell sample to flow through the filtering container for a filtering operation, and the cell sample flowing out of the filtering container will flow into the sorting container 120 for a fine sorting operation. In some embodiments, the control component can stop performing the filtering operation based on the second bubble sensor detecting a trigger stop signal. For more information about the filtering operation, please refer to the above description (for example, Figure 5C and step 2), which will not be repeated here.

[0117] Step 630: Use multiple sorting containers arranged in parallel to perform alternating sorting or simultaneous sorting.

[0118] In some embodiments, the sorting containers arranged in parallel can be multiple sorting containers 120 in Figures 1-3 and Figures 5A-5C. In some embodiments, alternating sorting can be that the sorting operations of multiple sorting containers are performed alternately. For example, at the current moment, at least one sorting container is sorting, and at least one sorting container is not sorting. After a period of time (such as 3 minutes), at least one sorting container is not sorting, and at least one sorting container is sorting. In some embodiments, simultaneous sorting can be that the sorting operations of multiple sorting containers are performed simultaneously. In some embodiments, the cell sorting device may include a second pipeline, and a first bubble sensor (for example, the first bubble sensor 181 in Figures 5A-5B) provided on the second pipeline. In some embodiments, the control component can stop alternating sorting or simultaneous sorting based on the first bubble sensor detecting a trigger stop signal. For more information about the subdividing sorting control operation, please refer to the above description (for example, Figures 5A-5C and step three), which will not be repeated here.

[0119] It should be noted that the above description of process 600 is for illustration and explanation only and does not limit the scope of application of this specification. For those skilled in the art, various modifications and changes can be made to process 600 under the guidance of this specification. However, these modifications and changes are still within the scope of this specification. For example, step 620 can be omitted, and the cells can be directly sorted alternately or simultaneously. For another example, step 630 can also include backflushing the cell transfer bag, flushing the sorting container and pipeline, releasing non-target cells adsorbed in the sorting container, or high-speed flushing of the coarse sorting container.

[0120] The beneficial effects that may be brought about by the embodiments of this specification include but are not limited to: (1) by setting up multiple sorting containers in parallel, multiple sorting containers can be alternately sorted or sorted simultaneously during the sorting process, increasing the sample processing capacity, shortening the sorting time, improving the sorting efficiency, reducing the reduction of cell activity and cell death caused by the long sorting time, breaking through the single sample sorting volume and sorting time limitations of existing equipment, and achieving high-quality cell sorting with large volume, high purity, high yield, reproducibility, and normal cell viability and cell function in a short time; (2) by combining the coarse sorting operation and the fine sorting operation, the purity of the sorted cells is higher, and the cell sorting efficiency is improved. It should be noted that different embodiments may have different beneficial effects. In different embodiments, the beneficial effects that may be produced may be any one or a combination of the above, or any other possible beneficial effects.

[0121] The basic concepts have been described above. It will be apparent to those skilled in the art that the detailed disclosure above is merely illustrative and does not limit the present application. Although not explicitly stated herein, those skilled in the art may make various modifications, improvements, and amendments to the present application. Such modifications, improvements, and amendments are suggested in the present application and remain within the spirit and scope of the exemplary embodiments of the present application.

[0122] At the same time, this application uses specific terms to describe the embodiments of this application. For example, "one embodiment," "an embodiment," and / or "some embodiments" refer to a certain feature, structure, or characteristic related to at least one embodiment of this application. Therefore, it should be emphasized and noted that "one embodiment," "an embodiment," or "an alternative embodiment" mentioned twice or multiple times in different locations in this specification does not necessarily refer to the same embodiment. In addition, certain features, structures, or characteristics in one or more embodiments of this application may be appropriately combined.

[0123] In addition, unless expressly stated in the claims, the order of the processing elements and sequences described in this application, the use of alphanumeric characters, or the use of other names are not intended to limit the order of the processes and methods of this application. Although the above disclosure discusses some of the invention embodiments currently considered useful through various examples, it should be understood that such details are only for illustrative purposes, and the attached claims are not limited to the disclosed embodiments. On the contrary, the claims are intended to cover all modifications and equivalent combinations that are consistent with the essence and scope of the embodiments of this application. For example, although the system components described above can be implemented by hardware devices, they can also be implemented only by software solutions, such as installing the described system on an existing server or mobile device.

[0124] Similarly, it should be noted that, in order to simplify the presentation of this application and thus facilitate understanding of one or more embodiments of the invention, the foregoing descriptions of the embodiments of this application sometimes combine multiple features into a single embodiment, figure, or description thereof. However, this disclosure method does not mean that the subject matter of this application requires more features than those recited in the claims. In fact, an embodiment may have fewer features than all of the features of a single embodiment disclosed above.

[0125] In some embodiments, numbers are used to describe the quantity of components and attributes. It should be understood that such numbers used in the description of the embodiments are modified by the modifiers "about", "approximately" or "substantially" in some examples. Unless otherwise stated, "about", "approximately" or "substantially" indicate that the numbers are allowed to vary by ±20%. Accordingly, in some embodiments, the numerical parameters used in the description and claims are approximate values, which may change according to the required features of individual embodiments. In some embodiments, the numerical parameters should take into account the specified significant digits and adopt the general method of retaining digits. Although the numerical domains and parameters used to confirm the breadth of their range in some embodiments of the present application are approximate values, in specific embodiments, the settings of such numerical values ​​are as accurate as possible within the feasible range.

[0126] Each patent, patent application, patent application disclosure, and other materials, such as articles, books, specifications, publications, documents, etc., cited in this application is hereby incorporated by reference in its entirety. This includes application history documents that are inconsistent with or conflict with the content of this application, as well as documents (currently or subsequently attached to this application) that limit the broadest scope of the claims of this application. It should be noted that if the descriptions, definitions, and / or use of terms in the accompanying materials of this application are inconsistent or conflicting with the content of this application, the descriptions, definitions, and / or use of terms in this application shall prevail.

[0127] Finally, it should be understood that the embodiments described in this application are merely illustrative of the principles of the embodiments of this application. Other variations may also fall within the scope of this application. Therefore, by way of example and not limitation, alternative configurations of the embodiments of this application may be considered consistent with the teachings of this application. Accordingly, the embodiments of this application are not limited to the embodiments explicitly introduced and described in this application.

Claims

1. A piping system for cell sorting, characterized in that: The pipeline system includes: a magnetic field component and a plurality of sorting containers arranged in the magnetic field component; wherein, The multiple sorting containers are arranged in parallel.

2. The piping system according to claim 1, characterized in that: The pipeline system further includes a sample bag and parallel pipelines arranged between the sample bag and the plurality of sorting containers.

3. The piping system according to claim 1, characterized in that: The magnetic field assembly includes a plurality of magnetic field modules, wherein each of the plurality of sorting containers is respectively arranged in one of the magnetic field modules.

4. The piping system according to claim 1, characterized in that: The magnetic field assembly includes a magnetic field module, wherein the multiple sorting containers share one magnetic field module.

5. The piping system according to claim 1, characterized in that: The magnetic field assembly includes a plurality of magnetic field modules, wherein the number of the sorting containers is greater than the number of the magnetic field modules, and at least two of the sorting containers share one magnetic field module.

6. The piping system according to claim 1, characterized in that: The pipeline system further includes a coarse sorting container, which is arranged in series before the multiple sorting containers.

7. The piping system according to claim 6, characterized in that: The pipeline system also includes a cell transfer bag, a first pipeline and a second pipeline; wherein the two ends of the first pipeline are respectively connected to the outlet end of the coarse sorting container and the first interface of the cell transfer bag, and the second pipeline is arranged between the second interface of the cell transfer bag and the inlet ends of the multiple sorting containers.

8. The piping system according to claim 7, characterized in that: The pipeline system further includes a first bubble sensor, which is disposed on the second pipeline.

9. The piping system according to claim 6, characterized in that: The pipeline system includes a first parallel pipeline, a second parallel pipeline and a junction pipeline; wherein, one end of the first parallel pipeline and the second parallel pipeline is connected to the outlet end of the coarse sorting container, and the other end is connected to the junction pipeline; at least one sorting container is respectively provided on the first parallel pipeline and the second parallel pipeline.

10. The piping system according to claim 9, characterized in that: The pipeline system includes a third parallel pipeline, which is arranged in parallel with the first parallel pipeline and the second parallel pipeline. One end of the third parallel pipeline is connected to the outlet end of the rough sorting container, and the other end is connected to the intersection pipeline.

11. The piping system according to claim 9, characterized in that: A first branch is provided on the first parallel pipeline, and the first branch is provided in parallel with the at least one sorting container on the first parallel pipeline.

12. The piping system according to claim 9, characterized in that A second branch is provided on the second parallel pipeline, and the second branch is provided in parallel with the at least one sorting container on the second parallel pipeline.

13. The piping system according to claim 9, characterized in that The pipeline system further includes a third waste liquid bag and a fifth pipeline, and both ends of the fifth pipeline are respectively connected to the third waste liquid bag and the junction pipeline.

14. The piping system according to claim 9, wherein: The pipeline system also includes a first terminal pipeline, a second terminal pipeline, a first waste liquid bag and a target cell bag; wherein, the two ends of the first terminal pipeline are respectively connected to the intersection pipeline and the first waste liquid bag, and the two ends of the second terminal pipeline are respectively connected to the intersection pipeline and the target cell bag.

15. The piping system according to claim 14, characterized in that: The pipeline system further includes a buffer bag and a buffer pipeline; wherein two ends of the buffer pipeline are respectively connected to the junction pipeline and the buffer bag.

16. The piping system according to claim 6, characterized in that The pipeline system further includes a second waste liquid bag and a third pipeline; wherein both ends of the third pipeline are respectively connected to the coarse separation container and the second waste liquid bag.

17. The piping system according to claim 6, characterized in that The pipeline system further includes a sample bag, a fourth pipeline and a second bubble sensor, wherein two ends of the fourth pipeline are respectively connected to the sample bag and the coarse sorting container, and the second bubble sensor is arranged on the fourth pipeline.

18. The piping system according to claim 6 or 15, characterized in that: The pipeline system also includes multiple pumps; wherein each of the multiple pumps is respectively arranged corresponding to the rough sorting container and one of the sorting containers, and the pumping speed of the pump corresponding to the rough sorting container is greater than the pumping speed of the pump corresponding to the sorting container.

19. A cell sorting device, characterized in that: The cell sorting device comprises the piping system according to any one of claims 1 to 18.

20. A method for controlling a cell sorting system, characterized in that: The method comprises: The cell sorting device is started, and multiple sorting containers arranged in parallel are used for alternating sorting or simultaneous sorting.

21. The control method according to claim 20, characterized in that: The method further comprises: Before performing the alternating sorting or the simultaneous sorting, a rough sorting control operation or a filtering operation is performed using a rough sorting container.

22. The control method according to claim 21, characterized in that: The rough sorting control operation includes: Start the coarse separation magnetic field component; The cell sample is controlled to flow through the coarse separation container, and the target cells flowing out of the coarse separation container are collected in a cell transfer bag.

23. The control method according to claim 21, characterized in that: The cell sorting device includes a fourth pipeline, and a second bubble sensor disposed on the fourth pipeline; the method further includes: The coarse sorting control operation or the filtering operation is stopped based on the second air bubble sensor detecting a trigger stop signal.

24. The control method according to claim 20, characterized in that: The cell sorting device includes a second pipeline and a first bubble sensor disposed on the second pipeline; the method further includes: The alternating sorting or the simultaneous sorting is stopped based on the first bubble sensor detecting a trigger stop signal.

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