Device for sorting and enriching

Abstract

The utility model provides a device for sorting and enriching, which comprises more than one sorting and enriching module and a power module, the separation and enrichment module comprises a separation and enrichment unit and a separation and enrichment unit, n sorting chips; a fluid inlet; a plurality of fluid outlets; in different sorting and enriching modules, N is independently an integer greater than or equal to 1; one or more sample storage containers; one or more buffer solution storage containers; two or more collection containers; the power module can enable fluid in the sample storage container and the buffer solution storage container to flow into the collection container through the sorting and enriching unit, and fluid in the collection container flows back to the cavity connected with the collection container. By means of the device, the cells and / or the cell clusters can be screened for multiple times, so that the sorting effect is improved, the sizes of the cells and / or the cell clusters collected in the collecting containers fall into the target size range more, and follow-up detection is facilitated.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biomedical technology, in particular to a device for sorting and enriching. BACKGROUND

[0002] Cell aggregates form cell clusters, which can be formed by aggregation of cells of the same type or cells of different types. Different types of cells in the cell cluster can play different roles, thereby endowing the cell cluster with biological functions that individual cells do not have and enabling the cell cluster to complete biological processes that individual cells cannot complete. The size of the cell cluster is related to the number of cells and the types of cells.

[0003] Malignant tumors are a serious disease, and most tumor patients die of tumor metastasis. In the past, it was believed that tumors metastasize through single tumor cells that circulate through the blood or lymph to reach distal tissues and form metastatic tumors, and therefore various techniques for separating and detecting single circulating tumor cells from blood have been developed. A typical separation and detection technique for single circulating tumor cells is the CellSearch product of Johnson & Johnson. Recent studies have shown that tumors mainly metastasize through tumor cell clusters. A tumor cell cluster can contain tumor cells of different properties and non-tumor cells. The size of a tumor cell cluster also varies, and the content of tumor cell clusters in the blood circulation system is very low. The number of tumor cell clusters in the blood is related to the prognosis of the patient. Since tumor cell clusters in the blood circulation system are not easy to obtain, there are also technical solutions to obtain dispersed single cells by digesting tumor tissue, and then re-aggregating the single cells to form tumor cell clusters for scientific research and clinical detection. However, the tumor cell clusters formed by this technical solution have a large size difference, and tumor cell clusters containing a certain number of cells and having uniform size need to be sorted and / or enriched to obtain stable and accurate detection results.

[0004] At present, the methods for sorting and / or enriching cells are generally divided into two categories: one is sorting and / or enriching cells according to their physical properties (cell size, density, motility, electrical properties, etc.), and the other is sorting and / or enriching cells according to their biochemical properties (surface antigens, etc.). Methods for sorting and / or enriching cells according to their physical properties, such as filtering and separating cells according to their size by designing a screen with a certain pore size, or sorting and / or enriching cells according to their density by density gradient centrifugation. Methods for sorting and / or enriching cells according to their biochemical properties, such as using fluorescently labeled antibodies to bind to cells according to their different surface antigens, and sorting and / or enriching cells by flow cytometry.

[0005] The existing cell sorting and / or enrichment method is mainly applied to the sorting and / or enrichment of single cells, and the sorting and / or enrichment method and technology of cell clusters are seriously lacking. Due to the difference in the number and type of cells, the size range of cell clusters is wide, and the sorting and / or enrichment technology of single cells is not suitable for the sorting and / or enrichment of cell clusters. The existing cell screen sorting and / or enrichment method mainly uses a mesh polymer membrane for filtration, and is mainly used for filtering and removing solid tissue blocks after the solid tissue is cut and digested. The mesh polymer membrane used by the cell screen has a pore size of more than 40 μm, which is not suitable for sorting and / or enriching free cell clusters existing in the in vivo fluid sample. The mesh polymer membrane is easy to block, and it is difficult to process a large volume of samples. The cell clusters stuck in the mesh polymer membrane are also difficult to recover, resulting in a large loss and being unable to be used for the separation of rare circulating tumor cell clusters. Practical new type content

[0006] To solve the problems in the prior art, the application provides a device for sorting and enrichment. The technical scheme of the application is as follows:

[0007] A device for sorting and enrichment, wherein,

[0008] The device comprises one or more sorting and enrichment modules and a power module.

[0009] The sorting and enrichment module comprises:

[0010] A sorting and enrichment unit, comprising: a shell; N sorting chips, the sorting chips being provided with a screening hole, and the N sorting chips being sequentially arranged to divide the internal space of the shell into a first cavity to an N+1 cavity; a fluid inlet provided on at least one side wall of the first cavity; a plurality of fluid outlets provided on the side walls of the first cavity to the N+1 cavity; and in different sorting and enrichment modules, N is an integer greater than or equal to 1.

[0011] One or more sample storage containers connected to the fluid inlet provided on the side wall of the first cavity through a flow channel;

[0012] One or more buffer solution storage containers connected to the fluid inlet through a flow channel;

[0013] Two or more collection containers connected to each fluid outlet through a flow channel;

[0014] The power module can make the fluid in the sample storage container and the buffer solution storage container flow into the collection container through the sorting and enrichment unit, and can make the fluid in the collection container connected to at least one of the first cavity to the N cavity flow back to the connected cavity.

[0015] Further, when N≥2, the fluid inlet is further arranged on the sidewall of at least one of the 2nd chamber to the N+1th chamber; and the fluid inlet is connected to the buffer storage container through a flow channel, respectively.

[0016] Further, when N≥2, the pore size of the screening hole of the sorting chip gradually decreases from the 1st chamber to the N+1th chamber.

[0017] Further, a control valve is arranged on the flow channel connecting the sample storage container and the fluid inlet on the sidewall of the 1st chamber; and / or, a control valve is arranged on at least one flow channel connecting the buffer storage container and the fluid inlet; and / or, a control valve is arranged on at least one flow channel connecting the collection container and the fluid outlet.

[0018] Further, the power module can control the positive and negative pressure in the collection container; and / or, the power module can control the positive and negative pressure in the sample storage container and the buffer storage container; and / or, the power module can provide positive pressure to the collection container, the sample storage container and the buffer storage container; and / or, the power module can provide negative pressure to the collection container, the sample storage container and the buffer storage container.

[0019] Further, in at least one of the sorting enrichment modules, the power module can independently control the pressure in the collection container, the sample storage container and / or the buffer storage container, respectively.

[0020] Further, at least one of the flow channels connecting the collection container and the fluid outlet is provided with a photoelectric detection unit and / or a flow detection unit.

[0021] Further, the photoelectric detection unit can detect absorbance and / or fluorescence.

[0022] Further, the device further comprises a pressure detection unit, which can detect the output pressure of the power module.

[0023] Further, at least one of the flow channels is a pipeline; preferably, the pipeline is a hose; further preferably, the control valve controls the opening and closing of the flow channel by extruding the hose from the outside.

[0024] Further, in at least one of the sorting and enriching modules, for at least one of the first chamber to the Nth chamber, the collection container connected to the chamber is directly or indirectly connected to the fluid inlet provided on the side wall of the chamber through a flow channel, the flow channel port of the flow channel is located at the bottom of the collection container, and a control valve for controlling the on-off is provided on the flow channel.

[0025] Further, the device comprises two or more sorting and enriching modules; wherein at least one collection container in at least one of the sorting and enriching modules is directly or indirectly connected to the fluid inlet provided on the side wall of the first chamber in other sorting and enriching modules through a flow channel; and the flow channel port of the flow channel is located at the bottom of the collection container.

[0026] By using the device for sorting and enriching provided in the present application, firstly, the fluid sample stored in the sample storage container and the buffer stored in the buffer storage container can flow into the sorting and enriching unit from the fluid inlet, so that the different sizes of cells and / or cell clusters after screening can enter the respective collection containers through the corresponding fluid outlet, and the cells and / or cell clusters in the corresponding size range can be screened and enriched; then, the fluid sample in the collection containers except the one connected to the N+1th chamber can flow into the sorting and enriching unit from the fluid outlet, and the screened cells and / or cell clusters can be screened again in the sorting chip. The device provided in the present application can repeatedly perform the above steps, so that the cells and / or cell clusters can be screened multiple times to improve the sorting effect, and the size of the cells and / or cell clusters collected in each collection container can fall into the target size range, thereby facilitating subsequent detection.

[0027] The above description is only a summary of the technical scheme of the present application. In order to make the technical means of the present application more clear and understandable, and to enable the person skilled in the art to implement the content of the description, and to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the specific embodiments of the present application are described below. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 : First step schematic diagram of the device for sorting and enriching in the first embodiment;

[0029] Figure 2 : Second step schematic diagram of the device for sorting and enriching in the first embodiment;

[0030] Figure 3 : First step schematic diagram of the device for sorting and enriching in the second embodiment;

[0031] Figure 4: Schematic diagram of the second step of the device for sorting and enriching in the second embodiment;

[0032] Figure 5 : Schematic diagram of the device for sorting and enriching in the third embodiment;

[0033] Figure 6 : Schematic diagram of the device for sorting and enriching in the fourth embodiment;

[0034] Figure 7 : Schematic diagram of the structure of the sorting and enriching unit;

[0035] Figure 8 : Schematic diagram of the structure of the upper shell;

[0036] Figure 9 : Schematic diagram of the structure of the lower shell;

[0037] Figure 10 : Schematic diagram of the structure of the sorting chip;

[0038] Figure 11 : Schematic diagram of the structure of the sorting chip in a wavy shape.

[0039] Explanation of reference numerals:

[0040] 110, sorting and enriching unit; 111, shell; 111-1, upper shell; 111-2, lower shell; 112, sorting chip; 113, screening hole; 114, fluid inlet; 115, fluid outlet; 116, micro column;

[0041] 120, sample storage container;

[0042] 130, buffer storage container;

[0043] 140, collection container;

[0044] V1~V8, control valve;

[0045] A1~A2, pump;

[0046] P1~P2, pressure detection unit;

[0047] F1~F4, photoelectric detection unit. DETAILED DESCRIPTION

[0048] The following embodiments of the present application are only used to illustrate the specific embodiments of the present application, and these embodiments cannot be understood as the limitation of the present application. Any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application are considered as equivalent replacement methods, which fall within the protection scope of the present application.

[0049] It should be understood by those skilled in the art that in the disclosure of the present application, the terms "first", "second", "third", "fourth", "fifth" and the like are only used to distinguish different structures, and do not limit the number of specific structures, connection relationship and the like; in addition, the orientation or position relationship indicated by "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer" and the like is based on the orientation or position relationship shown in the drawings, which is only for the convenience of describing the present application and simplifying the description, and does not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, therefore the above terms cannot be understood as a limitation of the present application.

[0050] The present embodiment provides a device for sorting and enriching, as shown in Figure 1 、 Figure 2 、 Figure 7 The device includes one or more (such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more) sorting and enrichment modules and a power module; wherein,

[0051] The sorting and enrichment module includes:

[0052] The sorting and enrichment unit 110 includes a shell 111, N sorting chips 112 provided with screening holes 113, and N sorting chips 112 arranged in sequence, which divide the internal space of the shell 111 into 1st cavity to N+1 cavity arranged in sequence; a fluid inlet 114 arranged at least on the side wall of the 1st cavity; a plurality of fluid outlets 115 arranged on the side wall of the 1st cavity to the N+1 cavity respectively; in different sorting and enrichment modules, N is an integer greater than or equal to 1 (such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more integers) independently;

[0053] One or more sample storage containers 120 are connected to the fluid inlet arranged on the side wall of the 1st cavity through a flow channel;

[0054] One or more buffer solution storage containers 130 are respectively connected to the fluid inlet through a flow channel;

[0055] Two or more collection containers 140 are respectively connected to each fluid outlet through a flow channel;

[0056] The power module enables the fluid in the sample storage container 120 and the buffer storage container 130 to flow into the collection container 140 through the sorting and enrichment unit, and enables the fluid in the collection container connected to at least one (such as one, part or all) of the first to Nth cavities to flow back to the cavity to which it is connected.

[0057] Those skilled in the art will know that when N=1, "at least one of the 1st cavity to the Nth cavity" refers to the 1st cavity.

[0058] Regarding the sorting and enrichment unit 110, the applicant has provided a sorting and enrichment device in Chinese patent applications CN202311586278.7 (Invention Title: A Device for Sorting and Enrichment and Its Application) and CN202323186250.2 (Utility Model Title: A Device for Sorting and Enrichment). This application further develops and applies the solution of that device (i.e., the sorting and enrichment unit indicated in this application) to increase sorting functionality and improve sorting capacity. The entire contents of both applications are incorporated herein by reference.

[0059] Specifically, such as Figure 7 As shown, the sorting and enrichment unit 110 includes: a housing 111; and N sorting chips 112 ( Figure 7 In the example, three sorting chips 112 are set. Each sorting chip 112 is provided with a screening hole 113. The N sorting chips 112 arranged sequentially divide the internal space of the housing 111 into the first cavity to the N+1th cavity arranged sequentially. Figure 7 In the example, the chambers are designated as 1st chamber, 2nd chamber, 3rd chamber, and 4th chamber from top to bottom; a fluid inlet 114 is disposed at least on the side wall of the 1st chamber; and multiple fluid outlets 115 are disposed on the side walls of the 1st chamber to the (N+1)th chamber, respectively. Figure 7 The example shows four fluid outlets (115).

[0060] This application does not specify a particular number of fluid inlets. Figures 1-6 As shown, only one fluid inlet is provided on the first cavity; as Figure 8 , Figure 10 As shown, the upper housing 111-1 can be combined with the sorting chip 112 to form two fluid inlets (and one fluid outlet) on the first cavity.

[0061] When there are two or more sorting and enriching modules, N is an integer greater than or equal to 1 in each of the different sorting and enriching modules, that is, the number of sorting chips of the sorting and enriching units in different sorting and enriching modules can be the same or different. Moreover, when there are two or more sorting and enriching modules, the pore sizes of the sorting chips of different sorting and enriching units can be the same or different.

[0062] In order to clearly and clearly illustrate the structure and working principle of the device for sorting and enriching of the present application, the embodiment mainly illustrates that the sorting and enriching unit only contains one sorting chip 112 (that is, N = 1).

[0063] The number and density of the screening holes 113 can be set as needed by those skilled in the art, and will not be repeated here. Specifically, a plurality of screening holes 113 are arranged in an array at a certain density.

[0064] The shape of the screening hole 113 is not specifically limited in the present application, as long as it can be applied to the screening of the corresponding cells and / or cell clusters. For example, the cross-sectional shape of the screening hole is circular or polygonal. In one sorting chip, the screening holes can be a combination of multiple shapes, that is, at least two of the screening holes have different shapes and / or pore sizes.

[0065] The pore size of each screening hole 113 can be set as needed by those skilled in the art. Specifically, in the present embodiment, the pore size of the screening hole is as small as 8 μm, that is, the pore size of the screening hole 113 in the present application is greater than or equal to 8 μm, specifically, 9 μm, 10 μm, 12 μm, 15 μm, 18 μm, 20 μm, 25 μm, 30 μm, 35 μm, 50 μm, 75 μm, 100 μm, 200 μm or 500 μm or more. Further, when the pore size of the screening hole 113 is less than or equal to 40 μm, the sorting chip is suitable for sorting and / or enriching free cell clusters present in the fluid sample. It should be noted that in the present application, the pore size refers to the diameter of the largest circle that can be accommodated by the cross section of the screening hole.

[0066] Preferably, a control valve is provided on the flow channel connecting the sample storage container 120 and the fluid inlet 114 on the side wall of the first cavity; and / or a control valve is provided on at least one (such as one, part or all) flow channel connecting the buffer storage container 130 and the fluid inlet 114; and / or a control valve is provided on at least one (such as one, part or all) flow channel connecting the collection container 140 and the fluid outlet 115. Specifically, the control valves V1~V6 in Figure 1 Figure 2 , so as to more accurately control the flow direction of the fluid and more accurately control the sorting and enrichment process. ​

[0067] As to the implementation of the power module, the present application is not particularly limited, as long as it can make the fluid in the sample storage container 120 and the buffer storage container 130 flow into the collection container 140 through the sorting enrichment unit, and can make the fluid in the collection container 140 connected to at least one (such as one, part or all) of the first cavity to the Nth cavity backflow to the cavity connected thereto. For example, the power module can control the pressure (such as gas pressure / liquid pressure) in the sample storage container 120, the buffer storage container 130 and / or the collection container 140, more specifically:

[0068] Option 1: The power module can control the positive and negative pressure (such as gas pressure / liquid pressure) in the collection container 140, so that the power module provides negative pressure to the collection container 140 to suck the fluid in the sample storage container 120 and the buffer storage container 130 to the collection container 140 through the sorting enrichment unit, and the power module can provide positive pressure to the collection container 140 to make the fluid in the collection container 140 backflow to the cavity connected thereto, at this time, it is preferred to set a control valve (such as control valve V1, control valve V2) that controls the opening and closing of the flow channel on the flow channel connected between the sample storage container 120, the buffer storage container 130 and the fluid inlet 114;

[0069] Option 2: The power module can control the positive and negative pressure (such as gas pressure / liquid pressure) in the sample storage container 120 and the buffer storage container 130, so that the power module provides positive pressure to the sample storage container 120 and the buffer storage container 130 to push the fluid in the sample storage container 120 and the buffer storage container 130 to the collection container 140 through the sorting enrichment unit, and the power module can provide negative pressure to the sample storage container 120 and the buffer storage container 130 to make the fluid in the collection container 140 backflow to the cavity connected thereto, at this time, it is preferred to set a control valve (such as control valve V3, control valve V4) that controls the opening and closing of the flow channel on the flow channel connected between the collection container 140 and the corresponding fluid outlet 115;

[0070] Option 3: The power module can provide positive pressure (such as gas pressure / liquid pressure) to the collection container, the sample storage container and the buffer storage container, so that the power module provides positive pressure to the sample storage container and the buffer storage container to push the fluid in the sample storage container and the buffer storage container to the collection container through the sorting enrichment unit, and the power module can provide positive pressure to the collection container to make the fluid in the collection container backflow to the cavity connected thereto;

[0071] Scheme 4, the power module can provide negative pressure (such as air pressure / hydraulic pressure) to the collection container 140, the sample storage container 120 and the buffer storage container 130, so as to provide negative pressure through the collection container 140 to suck the fluid in the sample storage container 120 and the buffer storage container 130 to the collection container 140 through the sorting enrichment unit, and can provide negative pressure to the sample storage container 120 and the buffer storage container 130 through the power module to return the fluid in the collection container to the cavity connected thereto.

[0072] In the following embodiments, the case where the power module adopts Scheme 1 (such as Figures 1-6 ) is mainly described.

[0073] In addition, as for the power source of the power module, when the power module provides hydraulic pressure (liquid pressure), a pump body such as a syringe pump can be listed; when the power module provides air pressure, a pump body (such as pump A1 and pump A2 in the drawings) such as an air pump and a peristaltic pump can be listed to provide positive pressure and / or negative pressure. In this application, it is preferred that the power module provides air pressure, and the power module can be connected to the collection container 140, the sample storage container 120 and / or the buffer storage container 130 through a flow channel to control the air pressure.

[0074] Preferably, in at least one (such as one, part or all) of the sorting enrichment modules, the power module can independently control the pressure (such as hydraulic pressure / air pressure) in the collection container, the sample storage container and / or the buffer storage container, respectively. Specifically as shown in Figures 1-2 , in this sorting enrichment module, different pumps are used to inflate or deflate different collection containers, so as to independently control the positive and negative pressure in each collection container.

[0075] Preferably, an air filtration unit is arranged at the inlet of the sample storage container 120, the inlet of the buffer storage container 130 and the air supply inlet of the power module to ensure a sterile environment in the device.

[0076] Preferably, a solid filtration unit (such as a filter screen) is arranged at the inlet of the flow channel through which the sample storage container 120 communicates with the cavity of the sorting enrichment unit 110 to prevent oversized solid substances in the sample from entering the flow channel or the sorting enrichment unit, thereby preventing the flow channel and / or the sorting chip sieve hole 113 from being blocked.

[0077] As for the material of the housing 111 and / or the sorting chip 112 in this application, it can be a metal material, an inorganic material and / or a polymer material, preferably an inorganic material (such as glass, silicon wafer, ceramic) and / or a polymer material (such as a polymer material).

[0078] The connection of the plurality of fluid inlets 114 to the buffer storage container 130 can be one-to-one, part-to-one, or all-to-one.

[0079] It should be noted that in the present application, the "cell cluster" refers to a cell aggregate formed by two or more cells combined together by covalent and / or non-covalent interaction, moving in a fluid as a whole.

[0080] When using the device of the present application for sorting and enrichment, the first step is to provide negative pressure to the pump A1 and the pump A2, as shown in the figure, so that the fluid sample stored in the sample storage container 120 and the buffer stored in the buffer storage container 130 flow into the sorting and enrichment unit 110 from the fluid inlet 114. When the fluid sample flows in the cavities, the flow direction is mainly divided into two directions: direction 1, for cells and / or cell clusters smaller than the pore size of the sorting chip 112, which pass through the sorting chip 112 and enter the lower cavity; direction 2, for cells and / or cell clusters larger than the pore size of the sorting chip 112, which cannot pass through the sorting chip 112 and can only flow to the fluid outlet 115 corresponding to the cavity. The specific flow direction is shown by the arrows, so that the sorted cells and / or cell clusters of different sizes enter the corresponding collection containers 140 through the corresponding fluid outlets 115, thereby realizing the sorting and enrichment of cells and / or cell clusters within a certain size range. Figure 1 The second step is to adjust the pump A1 to provide positive pressure and the pump A2 to provide negative pressure, as shown in the figure, so that the fluid sample collected in the collection container 140 connected to the first cavity flows into the sorting and enrichment unit 110 from the fluid outlet 115, and the already sorted cells and / or cell clusters are again subjected to sorting by the sorting chip 112, and the cells and / or cell clusters smaller than the sorting chip 112 flow into the second cavity and are collected in the collection container 140 connected to the second cavity. Figure 2 The second step is to adjust the pump A1 to provide positive pressure and the pump A2 to provide negative pressure, as shown in the figure, so that the fluid sample collected in the collection container 140 connected to the first cavity flows into the sorting and enrichment unit 110 from the fluid outlet 115, and the already sorted cells and / or cell clusters are again subjected to sorting by the sorting chip 112, and the cells and / or cell clusters smaller than the sorting chip 112 flow into the second cavity and are collected in the collection container 140 connected to the second cavity.

[0081] When N≥2, the pore size of the sorting chip decreases from the first cavity to the N+1 cavity. At this time, in the first step, two or more sorting chips 112 can sequentially sort cells and / or cell clusters, thereby sorting and collecting cells and / or cell clusters of multiple size ranges in the corresponding collection containers 140 in one step. In the second step, in addition to the collection container 140 connected to the N+1 cavity, the cells and / or cell clusters in the other collection containers 140 after the first step can be re-input into the sorting and enrichment unit 110 for re-sorting.

[0082] In addition, when the fluid inlets 114 are arranged on the side walls of at least two cavities in the sorting enrichment unit 110, the device of the present application can also achieve reverse flushing of the sorting chip 112 when the sorting chip 112 between the fluid inlets 114 is blocked, and the cells / cell clusters, etc. that block the screening holes 113 are flushed out of the screening holes 113 by the fluid. Specifically, taking the sorting enrichment unit 110 shown in the embodiments as an example, the fluid sample flows in from the fluid inlet 114 arranged on the side wall of the uppermost cavity of the first cavity (the first cavity) Figure 7 When the uppermost sorting chip (hereinafter also referred to as "the first sorting chip") is blocked, the power module of the above-mentioned scheme 1 can control the negative pressure in the uppermost collection container 140 to be the smallest, so that the buffer solution in the lower part of the first sorting chip flows from the lower part to the upper part of the sorting chip, thereby flushing the cells / cell clusters, etc. that block the screening holes 113 of the sorting chip back to the first cavity. In addition, the flow channel connecting the lower part of the first sorting chip and the corresponding cavity of the collection container 140 can also be closed (by controlling the valve, etc.), so that the buffer solution in the lower part of the first sorting chip flows from the lower part to the upper part of the sorting chip, thereby flushing the cells / cell clusters, etc. that block the screening holes 113 of the sorting chip back to the first cavity. Those skilled in the art know that for other sorting chips located between two fluid inlets 114 or other arrangements of the power module, the sorting chip can also be flushed in a similar manner. That is, when the fluid inlets 114 are arranged on the side walls of at least two cavities in the sorting enrichment unit 110, the pressure in the collection container 140 can be controlled and / or the flow channel connecting the collection container 140 and the corresponding cavity can be controlled (by controlling the valve, etc.) to open and close, so as to change the flow direction of the buffer solution in the sorting enrichment unit 110, and to flush the sorting chip between the fluid inlets 114 in the reverse direction to solve the problem of the sorting chip being blocked.

[0083] Therefore, the above-mentioned device of the present application can repeatedly cycle the above-mentioned first step and second step multiple times, so as to screen the cells and / or cell clusters multiple times to improve the sorting effect, so that the size of the cells and / or cell clusters collected in each collection container 140 falls more within the desired size range, thereby facilitating subsequent detection. Especially when the fluid inlets 114 are arranged on the side walls of at least two cavities in the sorting enrichment unit 110, the problem of the sorting chip being blocked can be solved.

[0084] Preferably, at least one (e.g. one, part or all) of the flow channels connecting the collection container 140 and the fluid outlet is provided with a photoelectric detection unit and / or a flow detection unit (in this embodiment, photoelectric detection unit F1 and photoelectric detection unit F2). The flow detection unit can detect the flow rate of the fluid, and the photoelectric detection unit (e.g. photoelectric detection unit F1 and photoelectric detection unit F2) can detect the absorbance and / or fluorescence of the cells / cell clusters (generally, the cells / cell clusters are sorted and enriched after being fluorescently labeled), so that the flow rate of the fluid and / or the flow rate of the cells / cell clusters in the pipeline can be accurately controlled by controlling the pressure, and / or the fluorescently labeled cells / cell clusters flow through the outlet, so that the sorting and enrichment process can be more accurately controlled.

[0085] Preferably, a pressure detection unit (e.g. pressure detection unit P1 and pressure detection unit P2, specifically a barometer) is provided in the flow channel connecting the power module and the collection container. The pressure detection unit can be used to conveniently control the pressure in the collection container, so as to control the flow rate and direction of the fluid in the flow channel, thereby more accurately controlling the sorting and enrichment process.

[0086] As for the flow channel in this application, it can be a pipeline or a channel provided in an entity. When it is a pipeline, it is preferably a flexible tube, so that the flexible tube can be squeezed from the outside by the control valve to control the opening and closing of the flow channel, without causing contamination of the fluid in the flexible tube.

[0087] In one embodiment of the above embodiment, as shown in Figure 7 when N≥2, the fluid inlet 114 is further provided on the side wall of at least one (e.g. one, part or all) of the 2nd chamber to the N+1th chamber, and preferably the fluid inlet 114 is further provided on the side wall of at least one (e.g. one, part or all) of the 2nd chamber to the Nth chamber; the fluid inlet 114 is connected to the buffer storage container 130 through a flow channel.

[0088] As known by those skilled in the art, when N=2, the "at least one of the 2nd chamber to the Nth chamber" refers to the 2nd chamber.

[0089] By providing a fluid inlet on the side wall of the corresponding chamber, the flow of the fluid sample in the corresponding chamber can be better driven by the buffer in the buffer storage container 130, and the cells and / or cell clusters can be reduced / prevented from blocking the screening hole 113.

[0090] In one embodiment of the above embodiment, the device further comprises a control module, which is electrically connected to the flow detection unit, the photoelectric detection unit, the pressure detection unit, the control valve and / or the power module.

[0091] As to the control module, it can specifically include a single-chip microcomputer, a PLC control machine, etc., to control the flow detection unit, the photoelectric detection unit, the pressure detection unit, and / or the control valve and / or the power module, so as to facilitate the automatic control of the device.

[0092] As to the control module, it can specifically include a single-chip microcomputer, a PLC control machine, etc., to control the flow detection unit, the photoelectric detection unit, the pressure detection unit, and / or the control valve and / or the power module, so as to facilitate the automatic control of the device.

[0093] In one embodiment of the above embodiment, as shown in Figures 8-10 The device further includes micro columns 116 (also referred to as "protrusions") in at least one (e.g., one, part or all) of the first to Nth cavities, and at least one (e.g., 1, 5, 10 or more than 20) micro columns 116 are arranged in the at least one cavity, and more preferably, the micro columns 116 are arranged in all the cavities.

[0094] Specifically, the micro columns 116 can be arranged on the shell (e.g., the upper shell 111-1, the lower shell 111-2, etc.) and / or the sorting chip 112, so that the micro columns 116 extend to the cavities.

[0095] As to the number and density of the micro columns 116, those skilled in the art can arrange them according to the needs, and the present application will not be repeated here. Specifically, a plurality of micro columns 116 can be arranged in an array in at least one (e.g., one, part or all) of the first to Nth cavities.

[0096] As to the cross-sectional shape of the micro columns 116, it can be circular, elliptical and / or polygonal, etc. according to the needs. Figures 8-10 For example, the cross-section of the micro column 116 is hexagonal and trapezoidal.

[0097] By arranging the above-mentioned micro columns 116 in the cavities, the original flow direction of the fluid in the cavities can be changed, the chances of the cells and / or cell clusters in the fluid sample contacting the screening holes 113 of the sorting chip 112 are increased, and the cells and / or cell clusters with a size smaller than the screening holes 113 can flow through the screening holes 113 to the next cavity as much as possible, rather than directly flowing to the fluid outlet 115 corresponding to the cavity, thereby enhancing the screening and enrichment effect of the device.

[0098] In one embodiment of the above embodiment, as shown in Figure 11As shown, the sorting chip 112 sets the area of the screening hole 113 to form a screening area; at least one (such as one, part or all) of the surfaces of the screening area of at least one (such as one, part or all) of the sorting chips 112 is / are partially or entirely in a non-planar structure, preferably, at least one (such as one, part or all) of the surfaces of the screening area of at least one (such as one, part or all) of the sorting chips 112 facing the first cavity is / are partially or entirely in a non-planar structure, and more preferably, at least one (such as one, part or all) of the surfaces of the screening area of at least one (such as one, part or all) of the sorting chips 112 is / are partially or entirely in a non-planar structure. Figure 11 Exemplarily, an example is given in which both surfaces of the screening area are curved (specifically, in a wave-like structure).

[0099] By making the surface of the screening area facing the first cavity in a non-planar structure, the original flow direction of the fluid in the cavity can be changed, and the chance of the cells and / or cell clusters contacting the screening hole 113 of the sorting chip 112 is increased, so that the cells and / or cell clusters with a size smaller than the screening hole 113 can flow to the next cavity through the screening hole 113 as much as possible, instead of directly flowing to the fluid outlet 115 corresponding to the cavity, thereby enhancing the screening and enrichment effect of the device.

[0100] In one embodiment of the above embodiment, as shown in Figure 3 , Figure 4 In at least one (such as one, part or all) of the sorting and enrichment modules, for at least one (such as one, part or all) of the cavities from the first cavity to the Nth cavity, the collection container 140 connected to the cavity is connected to the fluid inlet arranged on the side wall of the cavity through a flow channel (also referred to as a "special flow channel" in the specification), the flow channel port of the flow channel (special flow channel) is located at the bottom of the collection container, and a control valve (such as control valve V7) for controlling the on-off of the flow channel (special flow channel) is arranged on the flow channel (special flow channel), and a control valve (such as control valve V3) for controlling the on-off of the flow channel connecting the cavity and the collection container is arranged on the flow channel, the flow channel port of the flow channel (special flow channel) is located at the bottom of the collection container, so that most or all of the liquid in the collection container flows out of the collection container through the flow channel (special flow channel). In addition, a photoelectric detection unit (photoelectric detection unit F3) and / or a flow detection unit can also be arranged on the special flow channel.

[0101] As known by those skilled in the art, when N = 1, "at least one cavity from the first cavity to the Nth cavity" refers to the first cavity.

[0102] Figure 3 , Figure 4In the illustrated device, only the uppermost sorting enrichment module is provided with a special flow channel (special flow channel) connecting the collection container 140 and its corresponding fluid inlet; in the use of the sorting enrichment module of the device, the first step is only different from the first step of the above embodiment in that the control valve V7 is closed, and the fluid flow direction is as shown in Figure 3 ; the second step is only different from the second step of the above embodiment in that the control valve V7 is opened and the control valve V3 is closed, at which time the fluid sample collected in the collection container 140 connected to the first cavity flows into the sorting enrichment unit 110 from the specially provided flow channel (special flow channel), and the fluid flow direction is as shown in Figure 4 .

[0103] That is, the device provided by the present embodiment further increases a way for the fluid sample collected in the collection container 140 to flow into the sorting enrichment unit 110 on the basis of the above embodiment device. Those skilled in the art know that the first step and the second step can be repeated multiple times, so that the cells and / or cell clusters can be screened multiple times to improve the sorting effect, so that the size of the cells and / or cell clusters collected in each collection container 140 falls more within the desired size range, thereby facilitating subsequent detection.

[0104] In one embodiment of the above embodiment, as shown in Figure 5 , Figure 6 , the device includes two or more sorting enrichment modules; at least one (such as one, part or all) of at least one (such as one, part or all) collection container in at least one (such as one, part or all) of the sorting enrichment modules is connected to the fluid inlet provided on the side wall of the first cavity in the other sorting enrichment module through a flow channel (hereinafter also referred to as "external connection flow channel") (such as direct connection, or indirect connection to the fluid inlet 114 through the sample storage container 120 as in Figure 5 , Figure 6 ; and the flow channel port of the external connection flow channel is located at the bottom of the collection container 140, so that most or all of the liquid in the collection container flows out of the collection container through the external connection flow channel. In addition, the external connection flow channel can also be provided with a photoelectric detection unit (photoelectric detection unit F4) and / or a flow detection unit.

[0105] As shown in Figure 5 , Figure 6 , on the one hand, in one sorting enrichment module, negative pressure can be provided by the pump A1 and the pump A2 to screen the cells and / or cell clusters through the sorting enrichment unit 110 in the sorting enrichment module, and on the other hand, the fluid sample in the collection container 140 after screening is sucked into the sorting enrichment unit 110 in another sorting enrichment module, so as to perform re-screening in another sorting enrichment module. Therefore, the fluid sample can be screened multiple times through multiple sorting enrichment modules.

[0106] Preferably, when control valves (such as control valve V8, control valve V3) are arranged on the external flow channel, the collection container 140 connected to the corresponding fluid outlet, and the flow channel between them, then, as in the above embodiment, in one sorting enrichment module, the first step and the second step are only different from the above embodiment in that the control valve V8 is closed, so that the first step and the second step can be repeated in each sorting enrichment module to screen the cells and / or cell clusters multiple times. Then, through the external flow channel, enter another sorting enrichment module, and again be able to perform multiple screening, thereby further improving the sorting effect, so that the size of the cells and / or cell clusters collected in each collection container 140 falls more within the desired size range, thereby facilitating subsequent detection.

[0107] The device of any one of the above can be used to sort and enrich the following fluid samples:

[0108] (1) cells and / or cell clusters in peripheral blood samples;

[0109] (2) cells and / or cell clusters in pleural effusion, ascites effusion, lymph fluid, urine or cerebrospinal fluid;

[0110] (3) cells and / or cell clusters formed after enzymatic digestion of solid tissue; and / or,

[0111] (4) cell clusters formed by re-aggregation of single cells after digestion of solid tissue into single cells.

[0112] In addition, it can also be used to sort and enrich mixtures of particles of different sizes containing other biological molecules, such as (5) liposomes, water-in-oil microdroplets or oil-in-water microdroplets; and (6) other solid particulate matter, etc.

[0113] That is, in addition to providing the above-described device, the present application also provides the use of the above-described device in sorting and enriching the above-described fluid samples.

[0114] Although the embodiments of the present application are described above, the present application is not limited to the above-mentioned specific embodiments and application fields, and the above-mentioned specific embodiments are only illustrative and guiding, but not limiting. Those skilled in the art can make many forms under the guidance of the present specification and without departing from the scope protected by the claims of the present application, which all belong to the present application claimed.

Claims

1. An apparatus for sorting and enriching, wherein, Includes one or more sorting and enrichment modules and power modules; The sorting and enrichment module includes: A sorting and enrichment unit includes: a housing; N sorting chips, each sorting chip having a screening hole, the N sorting chips arranged sequentially dividing the internal space of the housing into sequentially arranged cavities from the first cavity to the (N+1)th cavity; a fluid inlet, which is at least located on the side wall of the first cavity; and multiple fluid outlets, which are respectively located on the side walls of the first cavity to the (N+1)th cavity; in different sorting and enrichment modules, N is an independent integer greater than or equal to 1. One or more sample storage containers are connected to a fluid inlet disposed on the side wall of the first cavity via a flow channel; One or more buffer storage containers are connected to the fluid inlet via flow channels; Two or more collection containers are connected to each of the fluid outlets via flow channels; The power module enables the fluid in the sample storage container and the buffer storage container to flow into the collection container through the sorting and enrichment unit, and enables the fluid in the collection container connected to at least one of the first to Nth cavities to flow back to the cavity to which it is connected.

2. The apparatus as claimed in claim 1, characterized in that, When N≥2, the fluid inlet is also provided on the side wall of at least one of the second to N+1 cavities; The fluid inlets are connected to the buffer storage container via flow channels.

3. The apparatus as described in claim 1, characterized in that, When N≥2, the aperture of the sorting chip gradually decreases from the first cavity to the N+1th cavity.

4. The apparatus as claimed in claim 1, characterized in that, A control valve is provided on the flow channel connecting the sample storage container and the fluid inlet on the side wall of the first cavity; and / or, A control valve is provided on at least one flow channel connecting the buffer storage container and the fluid inlet; and / or, A control valve is provided on at least one of the channels connecting the collection container and the fluid outlet.

5. The apparatus as claimed in claim 1, characterized in that, The power module can control the positive and negative pressure inside the collection container; and / or, The power module can control the positive and negative pressures within the sample storage container and the buffer storage container; and / or, The power module is capable of providing positive pressure to the collection container, the sample storage container, and the buffer storage container; and / or, The power module is capable of providing negative pressure to the collection container, the sample storage container, and the buffer storage container.

6. The apparatus as claimed in claim 1, characterized in that, In at least one of the sorting and enrichment modules, the power module is capable of independently controlling the pressure within the collection container, the sample storage container, and / or the buffer storage container.

7. The apparatus as claimed in claim 1, characterized in that, At least one of the flow channels connecting the collection container and the fluid outlet is provided with a photoelectric detection unit and / or a flow detection unit.

8. The apparatus as claimed in claim 7, characterized in that, The photoelectric detection unit can detect absorbance and / or fluorescence.

9. The apparatus as claimed in claim 1, characterized in that, The device also includes a pressure detection unit, which is capable of detecting the output pressure of the power module.

10. The apparatus as claimed in claim 4, characterized in that, At least one of the flow channels is a pipe.

11. The apparatus as claimed in claim 10, characterized in that, The pipeline is a flexible hose.

12. The apparatus as claimed in claim 11, characterized in that, The control valve controls the flow path by squeezing the hose from the outside.

13. The apparatus according to any one of claims 1 to 12, characterized in that, In at least one of the sorting and enrichment modules, for at least one of the first to Nth cavities, a collection container connected to the cavity is directly or indirectly connected to a fluid inlet disposed on the side wall of the cavity via a flow channel. The flow channel opening is located at the bottom of the collection container, and a control valve for controlling the on / off state is disposed on the flow channel. A control valve for controlling the on / off state is also disposed on the flow channel connecting the cavity and the collection container.

14. The apparatus according to any one of claims 1 to 12, characterized in that, The device includes two or more of the sorting and enrichment modules; In this embodiment, at least one collection container in at least one of the sorting and enrichment modules is directly or indirectly connected to a fluid inlet provided on the side wall of the first cavity in another sorting and enrichment module through a flow channel; and the flow channel opening is located at the bottom of the collection container.

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