Device for sorting and enrichment and use thereof

Abstract

Provided are a device for sorting and enrichment and the use thereof. The device comprises more than one sorting and enrichment module and a power module. The sorting and enrichment module comprises a sorting and enrichment unit, which comprises a housing, N sorting chips, a fluid inlet and a plurality of fluid outlets, N being independently an integer greater than or equal to 1 in different sorting and enrichment modules; more than one sample storage containers; more than one buffer solution storage containers; and more than two collection containers. The power module can enable a fluid in the sample storage containers and the buffer solution storage containers to flow through the sorting and enrichment unit into the collection containers, and the fluid in the collection containers to flow back to cavities connected thereto. The device can perform multiple times of screening on cells and / or cell clusters to improve the sorting effect, so as to enable the size of cells and / or cell clusters collected in each collection container to fall into a target size range, thereby facilitating subsequent tests.

Description

A device for sorting and enriching and its application Technical Field

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

[0002] Cell clusters can be formed by the aggregation of cells of the same or different cell types. Different cell types within a cell cluster may play different roles, enabling the cluster to possess biological functions not possessed by individual cells and to carry out biological processes that could not be accomplished by individual cells. The size of a cell cluster is related to the number and type of cells.

[0003] Malignant tumors are devastating diseases, with most cancer patients dying from metastasis. It was previously believed that tumors spread through single tumor cells that travel through the bloodstream or lymphatic system to distant tissues, forming metastases. Consequently, various technologies have been developed to isolate and detect single circulating tumor cells from the blood. Typical technologies for isolating and detecting single circulating tumor cells include Johnson & Johnson's CellSearch product. Recent studies have shown that tumors primarily metastasize through tumor cell clusters. Tumor cell clusters can contain tumor cells of varying natures as well as non-tumor cells. Tumor cell clusters vary in size and are present at very low levels in the bloodstream. The abundance of tumor cell clusters in the bloodstream correlates with patient prognosis. Because tumor cell clusters are difficult to obtain from the bloodstream, some approaches have employed digestion of tumor tissue to obtain dispersed single cells, which are then aggregated to form tumor cell clusters for research and clinical testing. However, the size of the tumor cell clusters generated by these techniques varies significantly, necessitating the separation and / or enrichment of tumor cell clusters containing a sufficient number of cells of uniform size to achieve stable and accurate detection results.

[0004] At present, cell sorting and / or enrichment methods are generally divided into two categories: one is sorting and / or enriching based on the physical properties of cells (cell size, density, motility, electrical properties, etc.), and the other is sorting and / or enriching based on the biochemical properties of cells (surface antigens, etc.). Methods for sorting and / or enriching based on the physical properties of cells include, for example, designing a sieve with a certain pore size for filtration separation based on the size of the cells, or sorting and / or enriching by density gradient centrifugation based on different cell densities. Methods for sorting and / or enriching based on the biochemical properties of cells include, for example, using fluorescently labeled antibodies to bind to cells based on different cell surface antigens, and sorting and / or enriching by flow cytometry.

[0005] Existing cell sorting and / or enrichment methods are mainly used for the sorting and / or enrichment of single cells, while there is a serious lack of methods and techniques for sorting and / or enriching cell clusters. Due to differences in cell numbers and cell types, the size range of cell clusters is wide, and single cell sorting and / or enrichment techniques are mostly not suitable for the sorting and / or enrichment of cell clusters. Existing cell sieve sorting and / or enrichment methods mainly use mesh polymer membranes for filtration, which are mostly used to remove solid tissue blocks after shearing and digestion of small-volume solid tissues. The pore size of the mesh polymer membranes used in cell sieves is mostly greater than 40μm, which is not suitable for sorting and / or enriching free cell clusters in intracellular fluid samples. Mesh polymer membranes are easily clogged and difficult to process large-volume samples. Cell clusters stuck in the mesh polymer membranes are also difficult to recover, resulting in large losses and cannot be used to separate rare circulating tumor cell clusters. Summary of the Invention

[0006] To solve the problems in the prior art, the present application provides a device for sorting and enriching, and the application of the device in sorting and enriching the following fluid samples. The technical solution of the present application is as follows:

[0007] 1. A device for sorting and enrichment, wherein:

[0008] Includes one or more sorting and enrichment modules and power modules;

[0009] The sorting and enrichment module comprises:

[0010] A sorting and enrichment unit comprises: a housing; N sorting chips, each having a screening hole, wherein the N sorting chips are sequentially arranged to divide the interior space of the housing into a first cavity to an N+1th cavity; a fluid inlet, which is at least provided on a side wall of the first cavity; and a plurality of fluid outlets, which are respectively provided on the side walls of the first cavity to the N+1th cavity; in different sorting and enrichment modules, N is independently an integer greater than or equal to 1.

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

[0012] one or more buffer storage containers connected to the fluid inlet via a flow channel;

[0013] Two or more collecting containers, each connected to the fluid outlet via a flow channel;

[0014] The power module can make the fluid in the sample storage container and the buffer 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 cavity from the first cavity to the Nth cavity flow back to the cavity to which it is connected.

[0015] 2. The device as described in item 1, wherein, when N≥2, the fluid inlet is also arranged on the side wall of at least one cavity from the second cavity to the N+1th cavity; the fluid inlet is connected to the buffer storage container through a flow channel.

[0016] 3. The device as described in item 1, wherein when N≥2, the aperture of the screening hole of the sorting chip gradually decreases from the 1st cavity to the (N+1)th cavity.

[0017] 4. The device as described in item 1, wherein 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 flow channel connecting the collection container and the fluid outlet.

[0018] 5. The device as described in item 1, wherein the power module is capable of controlling the positive and negative pressures in the collection container; and / or, the power module is capable of controlling the positive and negative pressures in 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.

[0019] 6. The device according to claim 1, wherein:

[0020] In at least one of the sorting and enrichment modules, the power module can independently control the pressures in the collection container, the sample storage container, and / or the buffer storage container.

[0021] 7. The device according to claim 1, wherein:

[0022] A photoelectric detection unit and / or a flow detection unit is provided on at least one of the flow channels respectively connecting the collection container and the fluid outlet.

[0023] 8. The device according to item 7, wherein:

[0024] The photoelectric detection unit is capable of detecting absorbance and / or fluorescence.

[0025] 9. The device according to claim 1, wherein:

[0026] The device further includes a pressure detection unit, which is capable of detecting the output pressure of the power module.

[0027] 10. The device according to claim 1, wherein:

[0028] At least one of the flow channels is a pipeline;

[0029] Preferably, the pipeline is a hose;

[0030] Further preferably, the control valve controls the on-off of the flow channel by squeezing the hose from the outside.

[0031] 11. The device according to any one of items 1 to 10, wherein

[0032] In at least one of the sorting and enrichment modules, for at least one cavity from the first cavity to the Nth cavity, the collection container connected to the cavity is directly or indirectly connected to the fluid inlet arranged on the side wall of the cavity through a flow channel, the flow channel mouth is located at the bottom of the collection container and a control valve for controlling on and off is arranged on the flow channel, and a control valve for controlling on and off is arranged on the flow channel connecting the cavity and the collection container.

[0033] 12. The device according to any one of items 1 to 10, wherein:

[0034] The device comprises two or more of the separation and enrichment modules;

[0035] Among them, 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 cavity in the other sorting and enriching modules through a flow channel; and the flow channel outlet is located at the bottom of the collection container.

[0036] 13. Use of the device according to any one of items 1 to 12 for sorting and enriching the following fluid samples:

[0037] cells and / or cell clusters in peripheral blood samples;

[0038] cells and / or cell clusters in pleural effusions, ascites, lymphatic fluid, urine, or cerebrospinal fluid;

[0039] Cells and / or cell clusters formed after enzymatic digestion of solid tissue;

[0040] Cell clusters formed by reaggregation of single cells after solid tissue is digested into single cells; or

[0041] Liposomes, water-in-oil microdroplets or oil-in-water microdroplets.

[0042] Sorting and enrichment are performed by the device for sorting and enrichment provided by the present application. First, the fluid sample stored in the sample storage container and the buffer stored in the buffer storage container can flow from the fluid inlet into the sorting and enrichment unit, so that the cells and / or cell clusters of different sizes after screening enter each collection container through the corresponding fluid outlet, respectively, to achieve screening and enrichment of cells and / or cell clusters in the corresponding size range; after that, the fluid samples in other collection containers except the collection container connected to the N+1 cavity can flow from the fluid outlet into the sorting and enrichment unit, and the screened ones enter the sorting chip again for screening. The above-mentioned device of the present application can screen cells and / or cell clusters multiple times by repeating the above steps multiple times to improve the sorting effect, so that the size of the cells and / or cell clusters collected in each collection container falls more into the target size range, which is beneficial to subsequent detection.

[0043] The above description is only an overview of the technical solution of the present application. In order to make the technical means of the present application clearer and easier to understand, so that those skilled in the art can implement it according to the contents of the specification, and to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are illustrated below. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1: Schematic diagram of the first step of the apparatus for sorting and enrichment in the first embodiment;

[0045] Figure 2: Schematic diagram of the second step of the device for sorting and enrichment in the first embodiment;

[0046] FIG3 is a schematic diagram of the first step of the apparatus for sorting and enrichment in the second embodiment;

[0047] FIG4 is a schematic diagram of the second step of the apparatus for sorting and enrichment in the second embodiment;

[0048] FIG5 is a schematic diagram of a device for sorting and enrichment in a third embodiment;

[0049] FIG6 is a schematic diagram of a device for sorting and enrichment in a fourth embodiment;

[0050] Figure 7: Schematic diagram of the structure of the sorting and enrichment unit;

[0051] Figure 8: Schematic diagram of the structure of the upper shell;

[0052] Figure 9: Schematic diagram of the structure of the lower shell;

[0053] Figure 10: Schematic diagram of the structure of the sorting chip;

[0054] Figure 11: Schematic diagram of the wavy structure of the sorting chip.

[0055] Description of the drawings: 110, sorting and enrichment unit; 111, shell; 111-1, upper shell; 111-2, lower shell; 112, sorting chip; 113, screening hole; 114, fluid inlet; 115, fluid outlet; 116, microcolumn; 120, sample storage container; 130, buffer storage container; 140, collection container; V1~V8, control valve; A1~A2, pump; P1~P2, pressure detection unit; F1~F4, photoelectric detection unit. DETAILED DESCRIPTION

[0056] The following embodiments of the present application are intended only to illustrate specific implementation methods for implementing the present application and are not to be construed as limiting the present application. Any other changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principles of the present application are deemed equivalent replacements and fall within the scope of protection of the present application.

[0057] Those skilled in the art should understand that, in the disclosure of this application, the terms "first", "second", "third", "fourth", "fifth", etc. are only used to distinguish different structures, and do not limit the number, connection relationship, etc. of specific structures; in addition, the orientations or positional relationships indicated by "longitudinal", "transverse", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. are based on the orientations or positional relationships shown in the accompanying drawings, which are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, the above terms cannot be understood as limitations on this application.

[0058] This embodiment provides a device for sorting and enrichment, as shown in Figures 1, 2, and 7, which includes one or more (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or more than 10) sorting and enrichment modules and a power module; wherein,

[0059] The sorting and enrichment module comprises:

[0060] The sorting and enrichment unit 110 includes: a housing 111; N sorting chips 112, each having a screening hole 113 provided thereon. The N sorting chips 112, which are arranged in sequence, divide the interior space of the housing 111 into a first cavity to an N+1th cavity, which are arranged in sequence; a fluid inlet 114, which is arranged at least on a side wall of the first cavity; and a plurality of fluid outlets 115, which are respectively arranged on the side walls of the first cavity to the N+1th cavity. In different sorting and enrichment modules, N is independently an integer greater than or equal to 1 (e.g., independently an integer greater than or equal to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10).

[0061] One or more sample storage containers 120 connected to the fluid inlet disposed on the side wall of the first cavity through a flow channel;

[0062] One or more buffer storage containers 130, which are respectively connected to the fluid inlet through flow channels;

[0063] Two or more collecting containers 140, which are respectively connected to the fluid outlets through flow channels;

[0064] The power module can allow 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 can allow the fluid in the collection container connected to at least one (such as one, part or all) of the 1st to Nth cavities to flow back to the cavity to which it is connected.

[0065] Those skilled in the art know that when N=1, “at least one cavity from the first cavity to the Nth cavity” refers to the first cavity.

[0066] Regarding the sorting and enrichment unit 110, the applicant provides a device for sorting and enrichment 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). Based on the scheme of the device (i.e., the sorting and enrichment unit indicated in this application), the present application further develops and applies it to increase the sorting function and improve the sorting capacity. The entire contents of the above two applications are incorporated into this application by reference.

[0067] Specifically, as shown in Figure 7, the sorting and enrichment unit 110 includes: a shell 111; N sorting chips 112 (in the example of Figure 7, three sorting chips 112 are set), and the sorting chip 112 is provided with a screening hole 113. The N sorting chips 112 arranged in sequence divide the internal space of the shell 111 into the first cavity to the N+1 cavity arranged in sequence (in the example of Figure 7, the first cavity, the second cavity, the third cavity, and the fourth cavity from top to bottom); a fluid inlet 114, which is at least arranged on the side wall of the first cavity; and multiple fluid outlets 115, which are respectively arranged on the side walls of the first cavity to the N+1 cavity (in the example of Figure 7, there are four fluid outlets 115).

[0068] This application does not specifically limit the number of fluid inlets. As shown in Figures 1 to 6 , only one fluid inlet is provided in the first cavity. As shown in Figures 8 and 10 , two fluid inlets (and one fluid outlet) can be formed in the first cavity by combining the upper housing 111 - 1 with the sorting chip 112 .

[0069] When there are two or more sorting and enrichment modules, N is independently an integer greater than or equal to 1 in each of the different sorting and enrichment modules. That is, the number of sorting chips in the sorting and enrichment units in different sorting and enrichment modules can be the same or different. Furthermore, when there are two or more sorting and enrichment modules, the apertures of the sorting chips in different sorting and enrichment units can be the same or different.

[0070] In order to clearly and concisely illustrate the structure and working principle of the device for sorting and enriching of the present application, the embodiments are mainly described as including only one sorting chip 112 (ie, N=1) in the sorting and enriching unit.

[0071] Regarding the number and density of the screening holes 113 , those skilled in the art can make specific settings as needed, and this application will not elaborate on them. Specifically, for example, a plurality of screening holes 113 are arranged in an array with a certain density.

[0072] Regarding the shape of the screening hole 113, the present application has no specific limitation, as long as it can be applied to the screening of corresponding cells and / or cell clusters. For example, the cross-sectional shape of the screening hole is circular or polygonal. In a sorting chip, the screening hole can be a combination of multiple shapes, that is, at least two of the screening holes have different shapes and / or pore sizes.

[0073] Regarding the pore size of each screening hole 113, those skilled in the art can make specific settings as needed. 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 8 μm or more, such as 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. Furthermore, 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.

[0074] Preferably, a control valve is provided on the flow channel connecting the sample storage container 120 and the fluid inlet 114 on the sidewall of the first chamber; and / or a control valve is provided on at least one (e.g., one, some, 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 (e.g., one, some, or all) flow channel connecting the collection container 140 and the fluid outlet 115. Specifically, control valves V1 to V6 are shown in Figures 1 and 2 , thereby more accurately controlling the flow direction of the fluid and thus more precisely controlling the sorting and enrichment processes.

[0075] The present application has no particular restrictions on the implementation of the power module, as long as it can allow 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 can allow the fluid in the collection container 140 connected to at least one (e.g., one, some, or all) of the first to Nth cavities to flow back to the connected cavity. For example, the power module can control the pressure (e.g., gas pressure / hydraulic pressure) within the sample storage container 120, the buffer storage container 130, and / or the collection container 140, more specifically, as follows:

[0076] Solution 1: The power module can control the positive and negative pressure (such as air pressure / hydraulic pressure) in the collection container 140, thereby providing negative pressure to the collection container 140 through the power module to draw the fluid in the sample storage container 120 and the buffer storage container 130 into the collection container 140 through the sorting and enrichment unit. The power module can also provide positive pressure to the collection container 140 to return the fluid in the collection container 140 to the connected cavity. In this case, control valves (such as control valve V1 and control valve V2) for controlling the flow channel connection between the sample storage container 120 and the buffer storage container 130 and the fluid inlet 114 are preferably provided.

[0077] Solution 2: The power module can control the positive and negative pressures (e.g., air pressure / hydraulic pressure) in the sample storage container 120 and the buffer storage container 130, thereby providing positive pressure to the sample storage container 120 and the buffer storage container 130 through the power module to push the fluid in the sample storage container 120 and the buffer storage container 130 to the collection container 140 through the sorting and enrichment unit, and can also 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 140 to the connected cavity. In this case, a control valve (e.g., control valve V3, control valve V4) for controlling the on-off of the flow channel connecting the collection container 140 and the corresponding fluid outlet 115 is preferably provided.

[0078] Solution 3: The power module is capable of providing positive pressure (such as air pressure / hydraulic pressure) to the collection container, the sample storage container, and the buffer storage container, thereby providing positive pressure to the sample storage container and the buffer storage container through the power module to push the fluid in the sample storage container and the buffer storage container through the sorting and enrichment unit to the collection container, and is capable of providing positive pressure to the collection container through the power module to return the fluid in the collection container to the cavity to which it is connected;

[0079] Option 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, thereby providing negative pressure through the collection container 140 to suck the fluid in the sample storage container 120 and the buffer storage container 130 into the collection container 140 through the sorting and 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 to which it is connected.

[0080] In the following embodiments, description is mainly made based on the case where the power module adopts solution 1 (such as the solutions shown in FIG. 1 to FIG. 6 ).

[0081] In addition, regarding the power source of the power module, when the power module provides hydraulic pressure (liquid pressure), a pump such as a syringe pump can be used; when the power module provides air pressure, a pump such as an air pump or a peristaltic pump can be used (such as pump A1 and pump A2 in the accompanying drawings) to provide positive and / or negative pressure. In the present application, it is preferred that the power module provides air pressure. For example, the power module can be connected to the collection container 140, the sample storage container 120, and / or the buffer storage container 130 via a flow channel to control the air pressure.

[0082] Preferably, in at least one (e.g., one, some, or all) of the sorting and enrichment modules, the power module is capable of independently controlling the pressure (e.g., hydraulic / pneumatic pressure) within the collection container, the sample storage container, and / or the buffer storage container. Specifically, as shown in Figures 1 and 2, in the sorting and enrichment module, different pumps are used to inflate or evacuate different collection containers, thereby independently controlling the positive and negative pressures within each collection container.

[0083] Preferably, air filtering units are provided 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 within the device.

[0084] Preferably, a solid filtering unit (such as a filter, etc.) is provided at the inlet of the flow channel connecting the sample storage container 120 and the cavity of the sorting and enrichment unit 110 to prevent solid matter of excessive size in the sample from entering the flow channel or the sorting and enrichment unit, thereby preventing blockage of the flow channel and / or the screening hole 113 of the sorting chip.

[0085] Regarding the material of the shell 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).

[0086] Regarding the connection method between the multiple fluid inlets 114 and the buffer storage container 130, each fluid inlet 114 can be connected to a buffer storage container 130, some of the fluid inlets 114 can be connected to a buffer storage container 130, or all of the fluid inlets can be connected to a buffer storage container 130.

[0087] It should be noted that, in the present application, "cell cluster" refers to a cell aggregate composed of two or more cells bound together by covalent and / or non-covalent interactions and moving in a fluid as a whole.

[0088] When using the device of the present application for sorting and enrichment, in the first step, as shown in Figure 1, pumps A1 and A2 both provide negative pressure, and the fluid sample stored in the sample storage container 120 and the buffer stored in the buffer storage container 130 flow from the fluid inlet 114 into the sorting and enrichment unit 110. When the fluid sample flows in each cavity, the flow direction is mainly divided into: movement direction 1, for cells and / or cell clusters with a smaller pore size than the screening hole 113 of the sorting chip 112, they pass through the screening hole 113 and enter the lower cavity; movement direction 2, for cells and / or cell clusters with a larger pore size than the screening hole 113 of the sorting chip 112, they cannot pass through the screening hole 113 of the sorting chip and can only flow to the fluid outlet 115 corresponding to the cavity in which they are located. The specific flow direction is shown by the arrows. As a result, the cells and / or cell clusters of different sizes after screening enter the collection containers 140 through the corresponding fluid outlets 115, thereby achieving screening and enrichment of cells and / or cell clusters of the corresponding size range. In the second step, as shown in Figure 2, pump A1 is adjusted to provide positive pressure and pump A2 to provide negative pressure. The specific flow direction of the fluid is shown by the arrow. The fluid sample collected and stored in the collection container 140 connected to the first cavity flows into the sorting and enrichment unit 110 from the fluid outlet 115. The screened fluid is screened again by the sorting chip 112. Cells and / or cell clusters smaller than the screening hole 113 are screened and flow into the second cavity and collected in the collection container 140 connected to the second cavity.

[0089] When N ≥ 2, the pore size of the screening holes of the sorting chip gradually decreases from the first cavity to the N+1th cavity. In this case, in the first step, two or more sorting chips 112 can screen cells and / or cell clusters step by step, thereby screening cells and / or cell clusters of various size ranges at once and collecting them in corresponding collection containers 140. In the second step, except for the collection container 140 connected to the N+1th cavity, the cells and / or cell clusters screened in the first step in the other collection containers 140 can be re-introduced into the sorting and enrichment unit 110 for further screening as described above.

[0090] In addition, preferably, when fluid inlets 114 are provided on the side walls of at least two cavities in the sorting and enrichment unit 110, when the sorting chip 112 between the fluid inlets 114 is blocked, the device of the present application can also realize reverse flushing of the sorting chip 112, and the cells / cell clusters that block the screening hole 113 are reversely flushed out of the screening hole 113 through the fluid. Specifically, taking the sorting and enrichment unit 110 shown in the embodiment as an example, its fluid sample flows into the fluid inlet 114 provided on the side wall of the first cavity (the uppermost cavity in FIG7 ). When the uppermost sorting chip (hereinafter also referred to as the "first sorting chip") is blocked, the power module using the above-mentioned scheme 1 can control the negative pressure in the uppermost collection container 140 to be minimum, so that the buffer solution at the bottom of the first sorting chip flows from the bottom to the top of the sorting chip, thereby flushing the cells / cell clusters blocking the screening holes 113 of the sorting chip back into the first cavity; in addition, the flow channel connecting the collection container 140 and the corresponding cavity at the bottom of the first sorting chip can be closed (by a control valve, etc.), so that the buffer solution flows from the bottom to the top of the first sorting chip, thereby flushing the cells / cell clusters blocking the screening holes 113 of the sorting chip back into the first cavity. Those skilled in the art will appreciate that for other sorting chips located between the two fluid inlets 114 or other arrangements of the power module, the sorting chip can also be backwashed in a similar manner. That is, when a fluid inlet 114 is provided on the side wall of at least two cavities in the sorting and enriching unit 110, the flow direction of the buffer solution in the sorting and enriching unit 110 can be changed by controlling the pressure in the collecting container 140 and / or controlling (controlling a valve, etc.) the on-off of the flow channel connecting the collecting container 140 and the corresponding cavity, and the sorting chip between the fluid inlet 114 can be backwashed to solve the problem of the sorting chip being blocked.

[0091] Therefore, the apparatus of the present application can repeatedly cycle through the first and second steps, thereby enabling multiple screening of cells and / or cell clusters, thereby improving the sorting effect and ensuring that the size of the cells and / or cell clusters collected in each collection container 140 falls more often within the target size range, thereby facilitating subsequent testing. In particular, when fluid inlets 114 are provided on the sidewalls of at least two cavities in the sorting and enrichment unit 110, the problem of clogging of the sorting chip can be resolved.

[0092] Preferably, at least one (such as one, some 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, the photoelectric detection unit F1 and the photoelectric detection unit F2). The flow detection unit can detect the flow rate of the fluid, and the photoelectric detection unit (such as the photoelectric detection unit F1 and the 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). This makes it possible to accurately control the fluid flow rate and / or the outflow rate of the cells / cell clusters in the pipeline and / or the outlet through which the fluorescently labeled cells / cell clusters flow by controlling the pressure, so as to more accurately control the sorting and enrichment process.

[0093] Preferably, a pressure detection unit (e.g., pressure detection unit P1, pressure detection unit P2, specifically, a barometer) is provided on the flow channel connecting the power module and the collection container. Providing a pressure detection unit can facilitate control of the pressure within the collection container, thereby controlling the flow rate and direction of the fluid in the flow channel, thereby more accurately controlling the sorting and enrichment process.

[0094] The flow channel in this application can be a pipeline or a channel provided in a physical body. When it is a pipeline, it is preferably a hose, so that the hose can be squeezed from the outside by a control valve to control the flow channel from opening to closing without contaminating the fluid in the hose.

[0095] In one implementation of the above embodiment, as shown in Figure 7, when N≥2, the fluid inlet 114 is also arranged on at least one (such as one, part or all) cavity side wall from the 2nd cavity to the N+1th cavity, and preferably, the fluid inlet 114 is also arranged on at least one (such as one, part or all) cavity side wall from the 2nd cavity to the Nth cavity; the fluid inlet 114 is connected to the buffer storage container 130 through a flow channel respectively.

[0096] Those skilled in the art know that when N=2, “at least one cavity from the second cavity to the Nth cavity” refers to the second cavity.

[0097] By arranging a fluid inlet on the side wall of the corresponding cavity, the buffer solution in the buffer solution storage container 130 can better drive the flow of the fluid sample in the corresponding cavity, thereby reducing / preventing cells and / or cell clusters from clogging the screening hole 113.

[0098] In one implementation of the above embodiment, the device further includes 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.

[0099] Regarding the control module, it may specifically include a single-chip microcomputer, a PLC controller, 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 to facilitate the automatic control of the device of this application.

[0100] Regarding the control module being electrically connected to the flow detection unit and / or the photoelectric detection unit to obtain corresponding signals, being electrically connected to the pressure detection unit to obtain pressure signals, being electrically connected to the control valve to control the action of the control valve, being electrically connected to the power module to control the gas flow direction and thus the positive and negative pressure, etc., all of these can be achieved through existing solutions and will not be elaborated here.

[0101] In one implementation of the above embodiment, as shown in Figures 8 to 10, the device further includes a microcolumn 116 (also referred to as a "protrusion"), and at least one (such as 1, 5, 10 or more than 20) microcolumns 116 are provided in at least one (such as one, part or all) of the 1st to Nth cavities. More preferably, the microcolumns 116 are provided in all of the above cavities.

[0102] Specifically, the microcolumns 116 may be disposed on a housing (such as the upper housing 111 - 1 , the lower housing 111 - 2 , etc.) and / or on the sorting chip 112 , so that the microcolumns 116 extend toward the cavity.

[0103] Regarding the number and density of each micropillar 116, those skilled in the art can make specific arrangements as needed, and this application will not elaborate on them in detail. Specifically, a plurality of micropillars 116 are arrayed in at least one cavity (e.g., one, some, or all) from the first cavity to the Nth cavity.

[0104] The cross-sectional shape of the micro-pillars 116 can be set to circular, elliptical and / or polygonal as required. Figures 8 to 10 exemplify examples of micro-pillars 116 having hexagonal and trapezoidal cross-sectional shapes.

[0105] By arranging the above-mentioned microcolumns 116 in the cavity, the original flow direction of the fluid in the cavity can be changed, thereby increasing the chances of cells and / or cell clusters in the fluid sample contacting the screening holes 113 of the sorting chip 112, so that cells and / or cell clusters 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 effects of the device of the present application.

[0106] In one implementation of the above embodiment, as shown in FIG11 , the area of ​​the sorting chip 112 where the screening holes 113 are provided forms a screening area; a portion or all of a surface of at least one (such as one, part, or all) of the screening area of ​​at least one (such as one, part, or all) of the sorting chip 112 is a non-planar structure, preferably, a portion or all of a surface of the screening area of ​​at least one (such as one, part, or all) of the sorting chip facing the first cavity is a non-planar structure, and further preferably, a portion or all of both surfaces of the screening area of ​​the sorting chip are non-planar structures. FIG11 exemplarily shows an example in which both surfaces of the screening area are curved surfaces (specifically, wavy structures).

[0107] By having a non-planar structure on the surface facing the first cavity in the screening area, the original flow direction of the fluid in the cavity can be changed, thereby increasing the chances of cells and / or cell clusters contacting the screening holes 113 of the sorting chip 112, so that cells and / or cell clusters 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 effects of the device of the present application.

[0108] In one implementation of the above embodiment, as shown in Figures 3 and 4, in at least one (e.g., one, some, or all) of the sorting and enrichment modules, for at least one (e.g., one, some, or all) of the cavities from the first cavity to the Nth cavity, the collection container 140 connected to the cavity is connected to a fluid inlet provided on the side wall of the cavity via a flow channel (also referred to as a "dedicated flow channel" in the specification). The flow channel (dedicated flow channel) has a flow channel opening located at the bottom of the collection container and is provided with a control valve (e.g., control valve V7) for controlling on / off. Furthermore, a control valve (e.g., control valve V3) for controlling on / off is provided on the flow channel connecting the cavity and the collection container. The flow channel (dedicated flow channel) has a flow channel opening 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 (dedicated flow channel). In addition, a photoelectric detection unit (photoelectric detection unit F3) and / or a flow detection unit may also be provided on the dedicated flow channel.

[0109] Those skilled in the art know that when N=1, “at least one cavity from the first cavity to the Nth cavity” refers to the first cavity.

[0110] In the devices illustrated in Figures 3 and 4, only the uppermost sorting and enriching module is provided with a special flow channel (dedicated flow channel) connecting the collection container 140 and its corresponding fluid inlet; when using the sorting and enriching module of the device, the first step differs from the first step of the above embodiment only in that the control valve V7 is closed, and the fluid flow direction is shown in Figure 3; the second step differs from the second step of the above embodiment only in that the control valve V7 is opened and the control valve V3 is closed. At this time, the fluid sample collected and stored in the collection container 140 connected to the first cavity flows from the specially provided flow channel (dedicated flow channel) into the sorting and enrichment unit 110, and the fluid flow direction is shown in Figure 4.

[0111] That is, the device provided in this embodiment further adds a method for the fluid sample collected in the collection container 140 to flow into the sorting and enrichment unit 110 based on the device of the above embodiment. Those skilled in the art will appreciate that the first and second steps can be performed multiple times, thereby enabling multiple screening of cells and / or cell clusters to improve the sorting effect and ensure that the size of the cells and / or cell clusters collected in each collection container 140 falls within the target size range, thereby facilitating subsequent testing.

[0112] In one implementation of the above embodiment, as shown in Figures 5 and 6, the device includes more than two of the aforementioned sorting and enrichment modules; wherein, at least one (such as one, some, or all) collection container in at least one (such as one, some, or all) of the aforementioned sorting and enrichment modules is connected to a fluid inlet provided on the side wall of the first cavity in another of the aforementioned sorting and enrichment modules via a flow channel (hereinafter also referred to as an "external flow channel") (specifically, such as direct connection, or, as shown in Figures 5 and 6, indirect connection to the fluid inlet 114 after passing through the sample storage container 120); and the flow channel opening of the flow channel is located at the bottom of the collection container 140, and the flow channel opening of the external 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 external flow channel. In addition, a photoelectric detection unit (photoelectric detection unit F4) and / or a flow detection unit may also be provided on the external flow channel.

[0113] As shown in Figures 5 and 6, in one sorting and enrichment module, negative pressure can be provided by both pumps A1 and A2 to screen cells and / or cell clusters through the sorting and enrichment unit 110 in that sorting and enrichment module. Furthermore, the screened fluid sample in the collection container 140 is drawn into the sorting and enrichment unit 110 in another sorting and enrichment module, where it is screened again. Therefore, multiple screening of the fluid sample can be performed using multiple sorting and enrichment modules.

[0114] Preferably, when control valves (e.g., control valve V8 or control valve V3) are provided on the flow channels between the external flow channel and the collection container 140 connected to the external flow channel and the corresponding fluid outlet, then, as in the above embodiment, in a sorting and enrichment module, the first and second steps differ from those in the above embodiment only in closing control valve V8. Thus, the first and second steps can be repeated multiple times in each sorting and enrichment module to screen cells and / or cell clusters multiple times. Thereafter, the cells can enter another sorting and enrichment module through the external flow channel and be screened multiple times again, thereby further improving the sorting effect and ensuring that the size of the cells and / or cell clusters collected in each collection container 140 falls more often within the target size range, thereby facilitating subsequent detection.

[0115] Any of the above devices can be used to sort and enrich the following fluid samples:

[0116] (1) Cells and / or cell clusters in peripheral blood samples;

[0117] (2) cells and / or cell clusters in pleural effusion, ascites, lymph, urine, or cerebrospinal fluid;

[0118] (3) cells and / or cell clusters formed after enzymatic digestion of solid tissue; and / or,

[0119] (4) Cell clusters formed by the reaggregation of single cells after solid tissue is digested into single cells.

[0120] 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, oil-in-water droplets or water-in-oil droplets; and (6) other solid particles.

[0121] That is, in addition to providing any of the above-mentioned devices, the present application also provides the application of the above-mentioned device in sorting and enriching the above-mentioned fluid samples.

[0122] 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. The above-mentioned specific embodiments are merely illustrative and instructive, and not restrictive. A person of ordinary skill in the art, guided by this specification and without departing from the scope of protection of the claims of this application, may also devise various forms, all of which fall within the scope of protection claimed in this application.

Claims

1. A device for sorting and enrichment, wherein: It includes one or more sorting and enrichment modules and power modules; The sorting and enrichment module comprises: The sorting and enrichment unit comprises: a housing; N sorting chips, each of which is provided with a screening hole, and the N sorting chips arranged in sequence divide the internal space of the housing into a first cavity to an N+1th cavity arranged in sequence; a fluid inlet, which is at least arranged on the side wall of the first cavity; and a plurality of fluid outlets, which are respectively arranged on the side walls of the first cavity to the N+1th cavity; in different sorting and enrichment modules, N is independently an integer greater than or equal to 1; One or more sample storage containers connected to the fluid inlet disposed on the side wall of the first cavity through a flow channel; One or more buffer storage containers connected to the fluid inlet via a flow channel; Two or more collecting containers, which are respectively connected to the fluid outlets through flow channels; The power module can make the fluid in the sample storage container and the buffer 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 cavity from the first cavity to the Nth cavity flow back to the cavity to which it is connected.

2. The device according to claim 1, wherein: When N≥2, the fluid inlet is also arranged on at least one side wall of the cavity from the second cavity to the N+1th cavity; The fluid inlets are connected to the buffer storage containers through flow channels, respectively.

3. The device according to claim 1, wherein: When N≥2, the aperture of the screening holes of the sorting chip gradually decreases from the first cavity to the N+1th cavity.

4. The device according to claim 1, wherein: 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 flow channel connecting the collecting container and the fluid outlet.

5. The device according to claim 1, wherein: The power module is capable of controlling the positive and negative pressures in the collection container; and / or, The power module is capable of controlling the positive and negative pressures in 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 device according to claim 1, wherein: In at least one of the sorting and enrichment modules, the power module can independently control the pressures in the collection container, the sample storage container and / or the buffer storage container.

7. The device of claim 1, wherein: A photoelectric detection unit and / or a flow detection unit is arranged on at least one of the flow channels respectively connecting the collection container and the fluid outlet.

8. The device according to claim 7, wherein: The photoelectric detection unit is capable of detecting absorbance and / or fluorescence.

9. The device of claim 1, wherein: The device further comprises a pressure detection unit, which is capable of detecting the output pressure of the power module.

10. The device of claim 1, wherein: At least one of the flow channels is a pipeline; Preferably, the pipeline is a hose; Further preferably, the control valve controls the on-off of the flow channel by squeezing the hose from the outside.

11. The device according to any one of claims 1 to 10, wherein: In at least one of the sorting and enrichment modules, for at least one cavity from the first cavity to the Nth cavity, a collecting container connected to the cavity is directly or indirectly connected to a fluid inlet arranged on the side wall of the cavity through a flow channel, a flow channel opening of the flow channel is located at the bottom of the collecting container and a control valve for controlling on and off is arranged on the flow channel, and a control valve for controlling on and off is arranged on the flow channel connecting the cavity and the collecting container.

12. The device according to any one of claims 1 to 10, wherein: The device comprises more than two of the sorting and enrichment modules; Among them, at least one collecting container in at least one of the sorting and enriching modules is directly or indirectly connected to the fluid inlet arranged on the side wall of the first cavity in other sorting and enriching modules through a flow channel; and the flow channel opening is located at the bottom of the collecting container.

13. Use of the device according to any one of claims 1 to 12 in sorting and enriching the following fluid samples: cells and / or cell clusters in peripheral blood samples; Cells and / or cell clusters in pleural effusions, ascitic effusions, lymphatic fluid, urine, or cerebrospinal fluid; Cells and / or cell clusters formed after enzymatic digestion of solid tissue; Cell clusters formed by reaggregation of single cells after solid tissues are digested into single cells; or Liposomes, water-in-oil microdroplets or oil-in-water microdroplets.

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