Device for sorting and enrichment and application thereof

Abstract

Provided are a device for sorting and enrichment and an application thereof. The device comprises a housing and N sorting chips, screening holes being formed in the sorting chips, and the N sequentially arranged sorting chips dividing the internal space of the housing into N+1 cavities, wherein N≥1. A first cavity is formed between one side of the housing and a sorting chip adjacent thereto; a second cavity is formed between the other side of the housing and a sorting chip adjacent thereto; a first outlet and a first inlet which connect the first cavity to the outside are formed in the cavity wall of the first cavity; and a second outlet which connects the second cavity to the outside is formed in the cavity wall of the second cavity.

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 and its application. Background Art

[0002] 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.

[0003] 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.

[0004] 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.

[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 circulating tumor cell clusters with rare content. Summary of the Invention

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

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

[0008] case;

[0009] N sorting chips, each having a screening hole, wherein the N sorting chips are sequentially arranged to divide the internal space of the housing into N+1 cavities, where N≥1;

[0010] in,

[0011] A first cavity is formed between one side of the housing and the adjacent sorting chip;

[0012] A second cavity is formed between the other side of the housing and the adjacent sorting chip;

[0013] A first outlet and a first inlet connecting the first cavity with the outside are formed on the cavity wall of the first cavity;

[0014] A second outlet connecting the second cavity and the outside is formed on the cavity wall of the second cavity.

[0015] 2. The device as described in item 1, wherein N≥2; a third cavity is formed between any adjacent sorting chips, and a third outlet connecting the third cavity with the outside is formed on the cavity wall of the third cavity.

[0016] 3. The device as described in item 2, wherein at least one of the third cavities has a third inlet formed on its cavity wall, connecting the third cavity with the outside world.

[0017] 4. The device as described in item 1, wherein two or more first inlets are formed on the cavity wall of the first cavity.

[0018] 5. The device as described in item 2, wherein the aperture of the screening holes of each of the sorting chips gradually decreases from one side of the shell to the other side of the shell; and / or the aperture of the screening holes is greater than or equal to 8 μm.

[0019] 6. The device as described in item 1, wherein the cross-sectional shape of the screening hole is circular or polygonal; and / or, in at least one of the sorting chips, at least two of the screening holes have different shapes.

[0020] 7. The device as described in item 2, wherein the device further comprises a microcolumn, and at least one microcolumn is arranged in at least one cavity among the first cavity, the second cavity, and the third cavity.

[0021] 8. The device as described in item 7, wherein the microcolumns are arranged on the shell; and / or the microcolumns are arranged on the surface of the sorting chip.

[0022] 9. The device as described in item 7, wherein the cross-section of the microcolumn is circular, elliptical or polygonal.

[0023] 10. The device as described in item 1, wherein the area where the screening holes are set in the sorting chip forms a screening area; a part or all of at least one surface of the screening area of ​​at least one of the sorting chips is a non-planar structure.

[0024] 11. The device as described in item 10, wherein a part or all of the surface of the screening area of ​​at least one of the sorting chips facing the first cavity is a non-planar structure.

[0025] 12. The device of item 10, wherein the non-planar structure is a curved surface.

[0026] 13. A device for sorting and enriching, comprising:

[0027] case;

[0028] N sorting chips, each having a screening hole, wherein the N sorting chips are sequentially arranged to divide the internal space of the housing into N+1 cavities, where N is greater than or equal to 2;

[0029] in,

[0030] A first cavity is formed between one side of the housing and the adjacent sorting chip;

[0031] A second cavity is formed between the other side of the housing and the adjacent sorting chip;

[0032] A third cavity is formed between any two adjacent sorting chips;

[0033] A first outlet connecting the first cavity and the outside is formed on the cavity wall of the first cavity;

[0034] A second outlet connecting the second cavity and the outside is formed on the cavity wall of the second cavity;

[0035] The third cavities are all provided with a third outlet connected to the third cavity and the outside world on their cavity walls, and at least one third cavity is provided with one or more third inlets connected to the third cavity and the outside world on its cavity wall.

[0036] 14. The device as described in item 13, wherein N=2; and / or at least one of the third cavities has two or more third inlets formed on its wall, connecting the third cavity with the outside.

[0037] 15. The device as described in item 14, wherein N=2; and the apertures of the screening holes 34b between the two sorting chips are the same.

[0038] 16. The device as described in item 13, wherein the pore size of the screening hole is greater than or equal to 8 μm.

[0039] 17. The device as described in item 13, wherein the cross-sectional shape of the screening hole is circular or polygonal; and / or, in at least one of the sorting chips, at least two of the screening holes have different shapes.

[0040] 18. The device as described in item 13, wherein the device further comprises a microcolumn, and at least one microcolumn is arranged in at least one cavity among the first cavity, the second cavity, and the third cavity.

[0041] 19. The device of item 18, wherein the microcolumns are arranged on the shell; and / or the microcolumns are arranged on the surface of the sorting chip.

[0042] 20. The device of item 19, wherein the cross-section of the microcolumns is circular, elliptical and / or polygonal.

[0043] 21. The device as described in item 13, wherein the area where the screening holes are set in the sorting chip forms a screening area; a part or all of at least one surface of the screening area of ​​at least one of the sorting chips is a non-planar structure.

[0044] 22. The device as described in item 21, wherein a part or all of a side of the screening area of ​​at least one of the sorting chips facing the third inlet is a non-planar structure.

[0045] 23. The device of item 21, wherein the non-planar structure is a curved surface.

[0046] 24. Use of the device described in any one of items 1 to 23 for sorting and enriching the following fluid samples: cell clusters in peripheral blood samples; cell clusters in pleural effusions, ascites effusions, lymphatic fluid, urine or cerebrospinal fluid; cell clusters formed after enzymatic digestion of solid tissues; cell clusters formed by reaggregation of single cells after solid tissues are digested into single cells; or liposomes, oil-in-water droplets or water-in-oil droplets.

[0047] The above-mentioned device for sorting and enrichment provided in the present application, through the following "movement direction 2", intercepts cells and / or cell clusters larger than the screening hole 34a in the corresponding cavity, and collects cells and / or cell clusters of the corresponding size range (larger size) through the corresponding outlet of the cavity, thereby screening and enriching the cells and / or cell clusters of the size range; through the following "movement direction 1", cells and / or cell clusters smaller than the screening hole pass through the screening hole into the lower cavity, thereby further achieving the screening and enrichment of cells and / or cell clusters of the corresponding size range (smaller size); when an inlet for providing a fluid such as a buffer is provided on the cavity, the fluid On the one hand, it can drive the flow of fluid samples in the corresponding cavity, reduce / prevent cells and / or cell clusters from clogging the screening holes, and at the same time, it can directly collect cells and / or cell clusters suspended in the corresponding fluid, thereby directly obtaining a suspension of cells and / or cell clusters that can be used for subsequent use; when the screening area of ​​the microcolumn and / or sorting chip is set to a non-planar surface, it can change the original flow velocity direction of the fluid in the cavity, increase the chance of cell clusters contacting the screening holes of the sorting chip, and make cell clusters smaller than the screening holes flow through the screening holes to the next cavity as much as possible, rather than directly flowing to the outlet corresponding to the cavity, thereby enhancing the screening and enrichment effect of the device of the present application.

[0048] 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

[0049] FIG1 is a schematic structural diagram of a device for sorting and enrichment in one embodiment of the present application;

[0050] Figure 2 is a schematic diagram of the inner structure of the upper shell in one embodiment of the present application;

[0051] Figure 3 is a schematic diagram of the outer structure of the upper shell in one embodiment of the present application;

[0052] FIG4 is a schematic diagram showing the upper structure of a first sorting chip in one embodiment of the present application;

[0053] FIG5 is a schematic diagram showing the lower structure of a first sorting chip in one embodiment of the present application;

[0054] Figure 6 is a schematic diagram of the inner structure of the lower housing in one embodiment of the present application;

[0055] Figure 7 is a schematic diagram of the outer structure of the lower housing in one embodiment of the present application;

[0056] FIG8 is a schematic diagram of the upper structure of the sorting chip not adjacent to the upper housing in one embodiment of the present application;

[0057] FIG9 is a schematic diagram of the lower structure of the sorting chip not adjacent to the upper housing in one embodiment of the present application;

[0058] FIG10 is a schematic diagram showing a structure in which the screening holes of the sorting chip are hexagonal in one embodiment of the present application;

[0059] FIG11 is a schematic diagram showing a structure in which the screening holes of a sorting chip are arranged in a combination of rectangular and circular shapes in one embodiment of the present application;

[0060] FIG12 is a schematic diagram showing a structure in which micropillars are arranged in an upper housing in one embodiment of the present application;

[0061] FIG13 is a schematic diagram showing a structure in which micropillars are arranged in the lower housing in one embodiment of the present application;

[0062] FIG14 is a schematic diagram showing a structure of micropillars provided on a sorting chip in one embodiment of the present application;

[0063] FIG15 is a schematic diagram showing a wavy structure of a sorting chip in one embodiment of the present application;

[0064] Figure 16 is a schematic structural diagram of a device for sorting and enrichment in another embodiment of the present application.

[0065] Explanation of the accompanying drawings: 10a, upper shell; 11a, upper shell side wall; 20a, lower shell; 21a, lower shell side wall; 30a, sorting chip; 31a, first sorting chip; 32a, second sorting chip; 33a, third sorting chip; 34a, screening hole; 35a, chip side wall; 41a, first cavity; 42a, second cavity; 43a, third cavity; 44a, first outlet; 45a, second outlet; 46a, third outlet; 47a, first inlet; 48a, third inlet; 50a, opening; 60a, microcolumn; 70a, cell cluster. 10b, upper shell; 20b, lower shell; 31b, first sorting chip; 32b, second sorting chip; 34b, screening hole; 41b, first cavity; 42b, second cavity; 43b, third cavity; 44b, first outlet; 45b, second outlet; 46b, third outlet; 48b, third inlet; 70b, cell cluster. DETAILED DESCRIPTION

[0066] 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.

[0067] This embodiment provides a device for sorting and enrichment, as shown in Figures 1 to 11, comprising:

[0068] case;

[0069] N sorting chips, each of which is provided with a screening hole 34a, are sequentially arranged to divide the internal space of the housing into N+1 cavities, where N≥1. As shown in FIG1 , in this embodiment, the N sorting chips are three sorting chips arranged from top to bottom—a first sorting chip 31a, a second sorting chip 32a, and a third sorting chip 33a; wherein,

[0070] A first cavity 41a is formed between one side of the housing (such as the inner side of the upper housing 10a) and the adjacent sorting chip (such as the first sorting chip 31a);

[0071] A second cavity 42a is formed between the other side of the housing (such as the inner side of the lower housing 20a) and the adjacent sorting chip (such as the third sorting chip 33a);

[0072] A first outlet 44a and a first inlet 47a are formed on the wall of the first cavity 41a, connecting the first cavity 41a with the outside.

[0073] A second outlet 45a connecting the second cavity 42a with the outside is formed on the cavity wall of the second cavity 42a.

[0074] There is no special limitation on the composition structure of the shell of the present application, as long as it can form an enclosed internal space. For example, it can be a separate closed shell with each sorting chip arranged in sequence inside it. It can also be composed of an upper shell 10a, a lower shell 20a and the chip side walls 35a of N sorting chips 30a arranged therebetween, as in the present embodiment.

[0075] Regarding the specific implementation method of the N sorting chips arranged in sequence to divide the internal space of the shell into N+1 cavities, the present application has no specific restrictions. For example, if N sorting chips are arranged in sequence in a closed shell, the N sorting chips divide the internal space of the shell into N+1 cavities. Alternatively, as in the present embodiment, the upper shell 10a, the sorting chip 30a and / or the lower shell 20a are concave, and N+1 cavities are formed by stacking the upper shell 10a, the lower shell 20a and the sorting chip 30a (refer to Figures 1 to 9).

[0076] Regarding the specific implementation of each outlet (such as the first outlet 44a, the second outlet 45a and the third outlet 46a described below) and / or inlet (the first inlet 47a and the second inlet and the third inlet 48a described below), the present application has no specific restrictions. For example, a channel connecting each cavity with the outside world is directly opened on the shell to obtain the corresponding inlet and / or outlet. Alternatively, as in the present embodiment, an opening 50a is provided on the side wall of the upper shell 10a, the sorting chip 30a and / or the lower shell 20a, and the corresponding inlet and / or outlet is obtained by stacking the upper shell 10a, the lower shell 20a and the sorting chip 30a (refer to Figures 1 to 9).

[0077] Regarding the number and density of the screening holes 34a, those skilled in the art can make specific settings as needed, and this application will not elaborate on them. Specifically, for example, multiple screening holes 34a are arranged in an array with a certain density.

[0078] Regarding the shape of the screening holes, this application has no specific restrictions, as long as it can be applied to the screening of corresponding cell clusters. For example, the cross-sectional shape of the screening holes is circular or polygonal. As shown in Figure 10, the cross-sectional shape of the screening holes is hexagonal, that is, an array or even multiple screening holes with hexagonal cross-sections. In a sorting chip, the screening holes can be a combination of multiple shapes. For example, in Figure 11, the screening holes are a combination of rectangular and circular. That is, in at least one (such as one, some or all) of the sorting chips, at least two screening holes have different shapes and / or pore sizes.

[0079] Regarding the pore size of each screening hole 34a, 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 8μm, that is, the pore size of the screening hole 34a in the present application is 8μm or more, such as 9μm, 10μm, 12μm, 15μm, 18μm, 20μm, 25μm, 30μm, 35μm, 50μm, 75μm, 100μm, 200μm or 500μm or more. Furthermore, when the pore size of the screening hole 34a is less than or equal to 40μm, 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.

[0080] The shells (such as the upper shell 10a and the lower shell 20a) and / or the sorting chip can be manufactured using existing and mature microfluidic chip processing technologies such as etching and injection molding. Subsequently, the complete sorting and enrichment device of the present application is formed through bonding and packaging.

[0081] Regarding the materials of the shell (such as the upper shell 10a, the lower shell 20a, etc.) and / or the sorting chip in this application, they can be metal materials, inorganic materials and / or polymer materials, preferably inorganic materials (such as glass, silicon wafers, ceramics) and / or polymer materials (such as polymer materials).

[0082] When using the device provided in this embodiment, a fluid sample is supplied from the first inlet 47a (here, a fluid sample containing a cell cluster is used as an example; the principle is the same for other fluid samples). By applying positive pressure to the first inlet 47a, applying negative pressure to the first outlet 44a, and / or applying negative pressure to the second outlet 45a, power can be provided for the flow of the fluid sample. When the fluid sample flows in each cavity (such as the first cavity 41a, the second cavity 42a, etc.), the flow direction is mainly divided into: movement direction 1, for cells and / or cell clusters with a smaller aperture than the screening hole 34a of the sorting chip, they pass through the screening hole 34a and enter the lower cavity; movement direction 2, for cells and / or cell clusters with a larger aperture than the screening hole 34a of the sorting chip, they cannot pass through the screening hole 34a of the sorting chip and can only flow to the outlet corresponding to the cavity where they are located. Then, as shown in Figure 1, the fluid sample is supplied from the first inlet 47a, and through the above-mentioned "movement direction 2", cells and / or cell clusters larger than the screening holes 34a of the first sorting chip 31a flow into the first outlet 44a, thereby screening and enriching cells and / or cell clusters in the corresponding size range (larger size); through the above-mentioned "movement direction 1", cells and / or cell clusters smaller than the screening holes 34a of the first sorting chip 31a pass through the screening holes 34a of the first sorting chip 31a into the lower cavity, thereby further realizing the screening and enrichment of cells and / or cell clusters in the corresponding size range (smaller size).

[0083] It should be noted that, in the present application, "cell cluster" refers to a cell cluster composed of two or more cells bound together by covalent and / or non-covalent interactions and moving in a fluid as a whole.

[0084] Preferably, when N≥2 (that is, when there are more than two sorting chips, such as 3, 4, 5, 6, 7, 8, 9 or more than 10, here taking the three sorting chips provided in this embodiment as an example), a third cavity 43a is formed between the sorting chips, and the third cavity 43a is formed with a third outlet 46a connecting the third cavity 43a with the outside world on its cavity wall. From one side of the shell (upper shell 10a) to the other side of the shell (lower shell 20a), the aperture of each screening hole of the sorting chip gradually decreases. At this time, it is also possible to choose to provide negative pressure through the third outlet 46a to provide power for the flow of the fluid sample. Then, the fluid sample flowing downward from the first cavity 41a into the third cavity 43a (here, the cavity between the first sorting chip 31a and the second sorting chip 32a) also has two flow directions. Among them, through the above-mentioned "movement direction 2", cells and / or cell clusters larger than the second sorting chip 32a flow into the third outlet 46a corresponding to the cavity, thereby screening and enriching cells and / or cell clusters in the corresponding size range; through the above-mentioned "movement direction 1", cells and / or cell clusters smaller than the second sorting chip 32a pass through the screening hole 34a of the second sorting chip 32a and enter the lower cavity, thereby screening and enriching cells and / or cell clusters in the corresponding size range. According to the same principle, from top to bottom, each sorting chip realizes the step-by-step screening of cells and / or cell clusters of different sizes and performs corresponding enrichment.

[0085] Preferably, more than two (two in this embodiment) first inlets 44a are formed on the wall of the first cavity 41a; and / or, at least one (such as one, part or all) of the third cavity is formed with a third inlet 48a connecting the third cavity 43a with the outside world on its wall. Generally, the cells and / or cell clusters after screening and enrichment need to be suspended in a corresponding fluid (such as a buffer and other components contained in the buffer) to obtain a suspension of cells and / or cell clusters for subsequent use. In this preferred embodiment, the corresponding fluid can be introduced through the extra first inlet 44a and / or third inlet 48a, and even positive pressure can be provided by the fluid. On the one hand, the introduced fluid can drive the flow of the fluid sample in the corresponding cavity, reduce / prevent cells and / or cell clusters from clogging the screening holes, and at the same time, the cells and / or cell clusters suspended in the corresponding fluid can be directly collected, thereby directly obtaining a suspension of cells and / or cell clusters that can be used for subsequent use.

[0086] Of course, a corresponding second inlet may also be provided on the second cavity 42a to facilitate the above-mentioned effect of reducing / preventing cells and / or cell clusters from clogging the screening holes and directly collecting cells and / or cell clusters suspended in the corresponding fluid.

[0087] In one implementation of the above embodiment, as shown in Figures 12 to 14, the device also includes a microcolumn 60a (also referred to as a "protrusion"), and at least one microcolumn 60a is arranged in at least one (such as one, part or all) of the first cavity 41a, the second cavity 42a, and the third cavity 43a. More preferably, the microcolumn 60a is arranged in all of the above cavities.

[0088] For example, the micropillars 60a can be arranged on the inner surface of the housing so that the micropillars 60a extend toward the first cavity 41a, the second cavity 42a, and / or the third cavity 43a. As shown in FIG12, a schematic diagram of the micropillars 60a arranged on the upper housing 10a is shown. The micropillars 60a are arranged on the inner side of the upper housing 10a, specifically, on the inner side of the upper housing sidewall 11a of the upper housing 10a and on the inner side of the upper housing 10a facing the adjacent sorting chip (the first sorting chip 31a). As shown in FIG13, a schematic diagram of the micropillars 60a arranged on the lower housing 20a is shown. The micropillars 60a are arranged on the inner side of the lower housing 20a, specifically, on the inner side of the lower housing sidewall 21a of the lower housing 20a and on the inner side of the lower housing 20a facing the adjacent sorting chip (the third sorting chip 33a). For another example, as shown in FIG14 , a schematic diagram of disposing a microcolumn 60 a on a sorting chip (specifically, a first sorting chip 31 a ) is shown. The microcolumn 60 a is disposed on the inner side of the chip sidewall 35 a .

[0089] The micropillars 60a can also be disposed on one side of the sorting chip facing the housing (e.g., toward the upper housing 10a) and / or the other side of the housing (e.g., toward the lower housing 20a). FIG. 14 shows a schematic diagram of micropillars 60a disposed on a sorting chip (specifically, the first sorting chip 31a). The micropillars 60a can be disposed on the upper side or, if desired, on the lower side. Specifically, the micropillars 60a can be disposed in the area of ​​the sorting chip where the screening holes 34a are disposed (i.e., the screening area described below).

[0090] The number and density of the micropillars 60a can be determined by those skilled in the art as needed, and will not be further described herein. Specifically, a plurality of micropillars 60a are arrayed in at least one (e.g., one, some, or all) of the first cavity 41a, the second cavity 42a, and the third cavity 43a.

[0091] The cross-sectional shape of the micro-pillars 60a can be set to circular, elliptical and / or polygonal as needed. Figures 12 to 14 exemplify examples of micro-pillars 60a having hexagonal and trapezoidal cross-sectional shapes.

[0092] By setting the above-mentioned microcolumns 60a in the cavity, the original flow direction of the fluid in the cavity can be changed, and the chances of cells and / or cell clusters 70a in the fluid sample contacting the screening holes 34a of the sorting chip 30a are increased, so that cell clusters smaller than the screening holes 34a can flow through the screening holes 34a to the next cavity as much as possible, rather than directly flowing to the outlet corresponding to the cavity, thereby enhancing the screening and enrichment effects of the device of the present application.

[0093] In one implementation of the above embodiment, the area where the screening holes 34a are set in the sorting chip 30a (e.g., the first sorting chip 31a, the second sorting chip 32a, and the third sorting chip 33a) forms a screening area; a portion or all of at least one surface of the screening area of ​​at least one (such as one, part, or all) of the sorting chips is a non-planar structure, preferably, a portion or all of the surface of the screening area of ​​at least one of the sorting chips 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. Figure 15 exemplifies an example in which both surfaces of the screening area are curved surfaces (specifically, wavy structures).

[0094] By providing 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 chance of the cell cluster 70a contacting the screening hole 34a of the sorting chip 30a, so that the cell clusters smaller than the screening hole 34a can flow through the screening hole 34a to the next cavity as much as possible, rather than directly flowing to the outlet corresponding to the cavity, thereby enhancing the screening and enrichment effects of the device of the present application.

[0095] In another embodiment of the present application, another device for sorting and enrichment is provided, as shown in FIG16 , comprising:

[0096] case;

[0097] N sorting chips, each of which is provided with a screening hole 34b, and the N sorting chips arranged in sequence divide the internal space of the housing into N+1 cavities, where N≥2. As shown in FIG16 , in this embodiment, the N sorting chips are two sorting chips arranged from top to bottom—a first sorting chip 31b and a second sorting chip 32b; wherein,

[0098] A first cavity 41b is formed between one side of the housing (such as the inner side of the upper housing 10b) and the adjacent sorting chip (ie, the first sorting chip 31b);

[0099] A second cavity 42b is formed between the other side of the housing (such as the inner side of the lower housing 20b) and the adjacent sorting chip (such as the second sorting chip 32b);

[0100] A third cavity 43b is formed between two adjacent sorting chips (such as between the first sorting chip 31b and the second sorting chip 32b);

[0101] A first outlet 44b is formed on the wall of the first cavity 41b, connecting the first cavity 41b with the outside;

[0102] A second outlet 45b is formed on the wall of the second cavity 42b, connecting the second cavity 42b with the outside.

[0103] The third cavities 43b are all formed with a third outlet 46b connecting the third cavity 43b with the outside world on their cavity walls, and at least one (such as one, part or all) of the third cavities 43b are formed with one or more third inlets 48b connecting the third cavity 43b with the outside world on their cavity walls.

[0104] In this embodiment, the composition structure of the shell; the specific implementation method of the N sorting chips arranged in sequence to divide the internal space of the shell into N+1 cavities; the specific implementation method of each outlet and / or inlet; the number and density of the screening holes; the shape and aperture of the screening holes; the preparation method and material of the shell and / or sorting chip; the setting position, size, density and shape of the microcolumns; the surface form of the screening area of ​​the sorting chip, etc., all refer to the previous embodiment, and those skilled in the art know that the same corresponding effects can be brought about, and will not be repeated here.

[0105] When using the device provided in this embodiment, a fluid sample is supplied through the third inlet 48b (here, a fluid sample containing a cell cluster is used as an example; the principle is the same for other fluid samples). By applying positive pressure at the third inlet 48b, negative pressure at the first outlet 44b, negative pressure at the second outlet 45b, and / or negative pressure at the third outlet 46b, power can be provided for the flow of the fluid sample. When the fluid sample flows in the third cavity 43b, the flow direction is mainly divided into: movement direction 1, for cells and / or cell clusters with a smaller aperture than the screening hole 34b of the sorting chip, they pass through the screening hole 34b and enter the adjacent cavity; movement direction 2, for cells and / or cell clusters with a larger aperture than the screening hole 34b of the sorting chip, they cannot pass through the screening hole 34b of the sorting chip and can only flow to the outlet corresponding to the cavity where they are located (the third outlet 46b). Then, as shown in FIG16 , the fluid sample is supplied from the third inlet 48 b, and through the above-mentioned “movement direction 2” in the third cavity 43 b, cells and / or cell clusters larger than the screening holes 34 b of the sorting chips on both sides of the third cavity 43 b (such as the first sorting chip 31 b and the second sorting chip 32 b) flow into the third outlet 46 b, thereby screening and enriching cells and / or cell clusters in the corresponding size range (larger size); through the above-mentioned “movement direction 1”, cells and / or cell clusters smaller than the screening holes 34 b of the sorting chips on both sides of the third cavity 43 b (such as the first sorting chip 31 b and the second sorting chip 32 b) pass through the screening holes 34 b of the first sorting chip 31 b or the second sorting chip 32 b and enter the cavities on both sides (such as the first cavity 41 b or the second cavity 42 b), thereby further screening and enriching cells and / or cell clusters in the corresponding size range (smaller size).

[0106] When the number of sorting chips on either side or both sides of the third inlet 48b for introducing fluid samples is greater than two, the aperture of the screening holes 34b between the sorting chips can be gradually reduced in the direction away from the third inlet 48b for introducing fluid samples, as in the previous embodiment. Thus, similar to the previous embodiment, cells and / or cell clusters of different sizes can be screened step by step and enriched accordingly.

[0107] Preferably, at least one of the third cavities 43b is formed with two or more third inlets 48b connecting the third cavity with the outside world on its wall. Generally, the cells and / or cell clusters after screening and enrichment need to be suspended in a corresponding fluid (such as a buffer solution and other costs contained in the buffer solution) to obtain a suspension of cells and / or cell clusters for subsequent use. In this preferred embodiment, one of the two or more third inlets 48b is used to introduce a fluid sample, and the extra third inlet 48b is used to introduce a corresponding fluid, and even provide positive pressure through the fluid. On the one hand, the fluid introduced can drive the flow of the fluid sample in the corresponding cavity, reduce / prevent cells and / or cell clusters from clogging the screening holes, and at the same time, cells and / or cell clusters suspended in the corresponding fluid can be directly collected, thereby directly obtaining a suspension of cells and / or cell clusters that can be used for subsequent use. Of course, in addition to setting two or more third inlets 48b on a third cavity 43b, corresponding inlets are set on other cavities (such as the first cavity 41b, the second cavity 42b and / or other third cavities 43b) (such as setting a first inlet on the first cavity; setting a second inlet on the second cavity; and / or, setting a third inlet in at least one (such as one, part or all) of the other third cavities), so that the corresponding fluid is introduced through the corresponding inlet, so as to achieve the above-mentioned effect of reducing / preventing cells and / or cell clusters from clogging the screening holes and directly collecting the cells and / or cell clusters suspended in the corresponding fluid.

[0108] Preferably, N = 2, and the filtration holes 34b of the sorting chips on both sides of the third cavity 43b (the first sorting chip 31b and the second sorting chip 32b) have the same pore size. In the fluid sample introduced through the third inlet 48b, cells and / or cell clusters smaller than the pore size of the filtration hole 34b enter the first and second cavities through the filtration hole 34b of the sorting chip, and flow out through the corresponding first and second outlets 44b and 45b and are collected. Cells and / or cell clusters larger than the pore size of the filtration hole 34b are retained by the sorting chip in the third cavity and flow out through the corresponding third outlet 46b and are collected. That is, after the cells and / or cell clusters in the fluid sample are sorted by the device, two cell and / or cell cluster components with different size ranges are obtained. When using the device to sort cells and / or cell clusters, the cells and / or cell clusters retained by the sorting chip can easily flow out of the device directly with the solvent through the third outlet, and are not likely to clog the screening holes 34b of the sorting chip, thereby improving the sorting and / or enrichment efficiency of the device; and the device has two sorting chips with the same pore size of the screening holes 34b, which is also conducive to improving the sorting and enrichment efficiency of the device, making the device suitable for processing large-volume samples.

[0109] Any of the above devices can be used to sort and enrich the following fluid samples:

[0110] (1) Cell clusters in peripheral blood samples;

[0111] (2) cell clusters in pleural effusion, ascites, lymph, urine, or cerebrospinal fluid;

[0112] (3) cell clusters formed after enzymatic digestion of solid tissue; and / or,

[0113] (4) Cell clusters formed by the reaggregation of single cells after solid tissue is digested into single cells.

[0114] 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.

[0115] 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.

[0116] 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 directions or positional relationships indicated by "longitudinal", "transverse", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. are based on the directions 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 direction, be constructed and operated in a specific direction. Therefore, the above terms cannot be understood as limiting this application.

[0117] 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: include: case; 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 N+1 cavities, where N≥1; in, A first cavity is formed between one side of the housing and the sorting chip adjacent thereto; A second cavity is formed between the other side of the housing and the sorting chip adjacent thereto; A first outlet and a first inlet connecting the first cavity with the outside are formed on the cavity wall of the first cavity; A second outlet connecting the second cavity and the outside is formed on the cavity wall of the second cavity.

2. The device according to claim 1, wherein: N≥2； A third cavity is formed between any adjacent sorting chips, and a third outlet connecting the third cavity with the outside is formed on the cavity wall of the third cavity.

3. The device as claimed in claim 2, wherein: At least one of the third cavities has a third inlet formed on its cavity wall, which connects the third cavity with the outside.

4. The device according to claim 1, wherein: More than two first inlets are formed on the cavity wall of the first cavity.

5. The device according to claim 2, wherein: The aperture of the screening holes of each of the sorting chips gradually decreases from one side of the shell to the other side of the shell; and / or, The pore size of the screening hole is greater than or equal to 8 μm.

6. The device according to claim 1, wherein: The cross-sectional shape of the screening hole is circular or polygonal; and / or, In at least one of the sorting chips, at least two of the screening holes have different shapes.

7. The device of claim 2, wherein: The device further comprises a microcolumn, and at least one microcolumn is disposed in at least one of the first cavity, the second cavity, and the third cavity.

8. The device according to claim 7, wherein: The microcolumn is arranged on the housing; and / or, The microcolumns are arranged on the surface of the sorting chip.

9. The device according to claim 7, wherein: The cross section of the microcolumn is circular, elliptical or polygonal.

10. The device of claim 1, wherein: The area where the screening holes are arranged on the sorting chip forms a screening area; A part or the whole of at least one surface of the screening area of ​​at least one of the sorting chips is a non-planar structure.

11. The device of claim 10, wherein: A part or the whole of a surface of the screening area of ​​at least one of the sorting chips facing the first cavity is a non-planar structure.

12. The device of claim 10, wherein: The non-planar structure is a curved surface.

13. A device for sorting and enrichment, wherein: include: case; 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 N+1 cavities, where N≥2; in, A first cavity is formed between one side of the housing and the sorting chip adjacent thereto; A second cavity is formed between the other side of the housing and the sorting chip adjacent thereto; A third cavity is formed between any two adjacent sorting chips; A first outlet connecting the first cavity with the outside is formed on the cavity wall of the first cavity; A second outlet connecting the second cavity and the outside is formed on the cavity wall of the second cavity; The third cavities are all provided with a third outlet connected to the third cavity and the outside world on the cavity wall thereof, and at least one third inlet connected to the third cavity and the outside world is formed on the cavity wall of at least one of the third cavities.

14. The device of claim 13, wherein: N=2; and / or, Two or more third inlets connecting the third cavity with the outside are formed on the cavity wall of at least one of the third cavities.

15. The device of claim 14, wherein: N=2; and the apertures of the screening holes 34b between the two sorting chips are the same.

16. The device of claim 13, wherein: The pore size of the screening hole is greater than or equal to 8 μm.

17. The device of claim 13, wherein: The cross-sectional shape of the screening hole is circular or polygonal; and / or, In at least one of the sorting chips, at least two of the screening holes have different shapes.

18. The device of claim 13, wherein: The device further comprises a microcolumn, and at least one microcolumn is disposed in at least one of the first cavity, the second cavity, and the third cavity.

19. The device of claim 18, wherein: The microcolumn is arranged on the housing; and / or, The microcolumns are arranged on the surface of the sorting chip.

20. The device of claim 19, wherein: The cross section of the microcolumn is circular, elliptical and / or polygonal.

21. The apparatus of claim 13, wherein: The area where the screening holes are arranged on the sorting chip forms a screening area; A part or the whole of at least one surface of the screening area of ​​at least one of the sorting chips is a non-planar structure.

22. The device of claim 21, wherein: A part or all of a surface of the screening area of ​​at least one of the sorting chips facing the third inlet is a non-planar structure.

23. The device of claim 21, wherein: The non-planar structure is a curved surface.

24. Use of the device according to any one of claims 1 to 23 in sorting and enriching the following fluid samples: Cell clusters in peripheral blood samples; Cell clusters in pleural effusions, ascites, lymphatic fluid, urine, or cerebrospinal fluid; 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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