Device for sorting and enriching

Abstract

The utility model provides a device for sorting and enriching, which comprises a shell, a separation device and an enrichment device, screening holes are formed in the sorting chips, the N sorting chips which are sequentially arranged divide the inner space of the shell into N + 1 cavities, and N is larger than or equal to 1; wherein a first cavity is formed between one side of the shell and the sorting chip adjacent to the shell; a second cavity is formed between the other side of the shell and the sorting chip adjacent to the shell; a first outlet and a first inlet which are used for connecting the first cavity with the outside are formed in the cavity wall of the first cavity; and a second outlet for connecting the second cavity with the outside is formed in the cavity wall of the second cavity.

Description

Technical Field

[0001] This application relates to the field of biomedical technology, specifically to a device for sorting and enrichment. Background Technology

[0002] Currently, cell sorting and / or enrichment methods are generally divided into two main categories: one based on the physical characteristics of cells (cell size, density, motility, electrical properties, etc.), and the other based on the biochemical characteristics of cells (surface antigens, etc.). Methods based on physical characteristics include, for example, using sieves with specific pore sizes for filtration based on cell size, or using density gradient centrifugation based on cell density. Methods based on biochemical characteristics include, for example, using fluorescently labeled antibodies to bind to cells based on different cell surface antigens, and then using flow cytometry for sorting and / or enrichment.

[0003] Cell clusters, formed by the aggregation of cells, may consist of cells of the same type or different types. Different cell types within a cluster may play different roles, giving the cluster biological functions that a single cell cannot perform and enabling biological processes that a single cell cannot complete. The size of a cell cluster is related to the number and type of cells.

[0004] Malignant tumors are a serious disease, and most cancer patients die from tumor metastasis. Past understanding held that tumors spread to distant tissues as individual tumor cells via the bloodstream or lymphatic system, forming metastases. Therefore, various techniques for isolating and detecting individual circulating tumor cells from the blood were developed. Typical techniques for isolating and detecting individual circulating tumor cells include Johnson & Johnson's CellSearch product. Recent studies have shown that tumors primarily metastasize through tumor cell clusters. Tumor cell clusters may contain tumor cells of different types, as well as non-tumor cells. The size of tumor cell clusters also varies, and their concentration in the bloodstream is very low. The amount of tumor cell clusters in the blood is related to patient prognosis. Because tumor cell clusters in the bloodstream are not easily obtained, some techniques involve digesting tumor tissue to obtain dispersed individual cells, which are then aggregated to form tumor cell clusters for research and clinical testing. However, tumor cell clusters formed using this technique vary significantly in size, requiring sorting and / or enrichment of tumor cell clusters containing a certain number of cells and of uniform size to obtain stable and accurate test results.

[0005] Existing cell sorting and / or enrichment methods are mainly applied to the sorting and / or enrichment of single cells, while methods and techniques for sorting and / or enriching cell clusters are severely lacking. Due to the differences in cell number and cell species, cell cluster sizes vary widely, making single-cell sorting and / or enrichment techniques largely unsuitable for cell cluster sorting and / or enrichment. Existing cell sieving and / or enrichment methods primarily use mesh polymer membranes for filtration, mostly used for filtering out small-volume solid tissues after mincing and digestion. The pore size of the mesh polymer membranes used in cell sieving is mostly greater than 40 μm, making them unsuitable for sorting and / or enriching free cell clusters present in fluid samples. Mesh polymer membranes are prone to clogging, making it difficult to process large-volume samples. Cell clusters trapped within the mesh polymer membrane are also difficult to recover, resulting in significant losses and making them unsuitable for separating sparse circulating tumor cell clusters. Utility Model Content

[0006] To address the problems in the prior art, this application provides an apparatus for sorting and enriching. The technical solution of this application is as follows:

[0007] An apparatus for sorting and enriching, comprising:

[0008] case;

[0009] N sorting chips, each with a screening hole, are arranged sequentially 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] The cavity wall of the first cavity has a first outlet and a first inlet that connect the first cavity to the outside.

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

[0015] Furthermore, N≥2; a third cavity is formed between any adjacent sorting chips, and a third outlet is formed on the cavity wall of each third cavity to connect the third cavity with the outside.

[0016] Furthermore, at least one of the third cavities has a third inlet formed on its cavity wall, connecting the third cavity to the outside.

[0017] Furthermore, two or more first inlets are formed on the cavity wall of the first cavity.

[0018] Furthermore, along the direction from one side of the housing to the other side, the aperture of the screening hole of each sorting chip gradually decreases; and / or, the aperture of the screening hole is greater than or equal to 8 μm.

[0019] Furthermore, 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] Furthermore, the device also includes micropillars, with at least one micropillar disposed in at least one of the first cavity, the second cavity, and the third cavity.

[0021] Furthermore, the micropillars are disposed on the housing; and / or, the micropillars are disposed on the surface of the sorting chip.

[0022] Furthermore, the cross-section of the micropillar is circular, elliptical, or polygonal.

[0023] Furthermore, the area where the screening holes are set on the sorting chip forms a screening area; at least a portion or all of at least one surface of the screening area of ​​at least one sorting chip is a non-planar structure.

[0024] Furthermore, at least one of the screening areas of the sorting chip facing the first cavity has a portion or all of a non-planar structure.

[0025] Furthermore, the non-planar structure is a curved surface.

[0026] This application provides another apparatus for sorting and enrichment, comprising:

[0027] case;

[0028] N sorting chips, each sorting chip having a screening hole, and the N sorting chips arranged sequentially divide the internal space of the housing into N+1 cavities, where N≥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 is formed on the cavity wall of the first cavity, connecting the first cavity to the outside.

[0034] A second outlet is formed on the cavity wall of the second cavity, connecting the second cavity to the outside.

[0035] Each of the third cavities has a third outlet on its cavity wall that connects the third cavity to the outside, and at least one of the third cavities has one or more third inlets on its cavity wall that connect the third cavity to the outside.

[0036] Furthermore, N=2; and / or, at least one of the third cavities has two or more third entrances formed on its cavity wall, connecting the third cavity to the outside.

[0037] Furthermore, N=2; and the aperture of the screening hole 34b is the same between the two sorting chips.

[0038] Furthermore, the pore size of the screening hole is greater than or equal to 8 μm.

[0039] Furthermore, 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] Furthermore, the device also includes micropillars, with at least one micropillar disposed in at least one of the first cavity, the second cavity, and the third cavity.

[0041] Furthermore, the micropillars are disposed on the housing; and / or, the micropillars are disposed on the surface of the sorting chip.

[0042] Furthermore, the cross-section of the micropillar is circular, elliptical, and / or polygonal.

[0043] Furthermore, the area where the screening holes are set on the sorting chip forms a screening area; at least a portion or all of at least one surface of the screening area of ​​at least one sorting chip is a non-planar structure.

[0044] Furthermore, at least one of the sorting chips has a non-planar structure on one or all of the side of the screening area facing the third inlet.

[0045] Furthermore, the non-planar structure is a curved surface.

[0046] The sorting and enrichment apparatus provided in this application, via "movement direction 2" as described below, traps cells and / or cell clusters larger than the screening well 34a within the corresponding cavity, and collects cells and / or cell clusters of a corresponding size range (larger size) through the corresponding outlet of the cavity, thereby screening and enriching cells and / or cell clusters of that size range; via "movement direction 1" as described below... Cells and / or cell clusters smaller than the screening wells are allowed to pass through the screening wells into the lower cavity, thereby further screening and enriching cells and / or cell clusters within the corresponding size range (smaller size). When an inlet for providing fluids such as buffer solutions is provided on the cavity, this fluid can, on the one hand, drive the flow of fluid samples within the corresponding cavity, reducing / preventing cell and / or cell cluster blockage of the screening wells. On the other hand, it can directly collect cells and / or cell clusters suspended in the corresponding fluid, thus directly obtaining a suspension of cells and / or cell clusters that can be used subsequently. When the screening area of ​​the microcolumn and / or the sorting chip is set to be non-planar, it can change the original flow direction of the fluid within the cavity, increasing the chance of cell clusters contacting the screening wells of the sorting chip. This allows cell clusters smaller than the screening wells to flow through the screening wells to the next cavity as much as possible, rather than flowing directly to the outlet corresponding to their current cavity, thereby enhancing the screening and enrichment function of the device in this application.

[0047] The above description is merely an overview of the technical solution of this application. In order to make the technical means of this application clearer and more understandable, so that those skilled in the art can implement it according to the contents of the specification, and in order to make the above and other objects, features and advantages of this application more obvious and understandable, the following is an example of a specific implementation of this application. Attached Figure Description

[0048] Figure 1 : A schematic diagram of the structure of an apparatus for sorting and enrichment in one embodiment of this application;

[0049] Figure 2 : A schematic diagram of the inner structure of the upper shell in one embodiment of this application;

[0050] Figure 3 : A schematic diagram of the outer structure of the upper shell in one embodiment of this application;

[0051] Figure 4 : A schematic diagram of the upper structure of the first sorting chip in one embodiment of this application;

[0052] Figure 5 A schematic diagram of the lower structure of the first sorting chip in one embodiment of this application;

[0053] Figure 6 : A schematic diagram of the inner structure of the lower housing in one embodiment of this application;

[0054] Figure 7 : A schematic diagram of the outer structure of the lower housing in one embodiment of this application;

[0055] Figure 8 : A schematic diagram of the upper structure of the sorting chip that is not adjacent to the upper housing in one embodiment of this application;

[0056] Figure 9 : A schematic diagram of the lower structure of the sorting chip that is not adjacent to the upper housing in one embodiment of this application;

[0057] Figure 10 : A schematic diagram of the sorting chip's hexagonal screening holes in one embodiment of this application;

[0058] Figure 11 : A schematic diagram of a structure in one embodiment of this application showing that the sorting chip has a combination of rectangular and circular screening holes;

[0059] Figure 12 A schematic diagram of a structure in one embodiment of this application, showing a micropillar disposed inside the upper shell;

[0060] Figure 13 A schematic diagram of a structure in which micropillars are disposed inside the lower shell in one embodiment of this application;

[0061] Figure 14 A schematic diagram of a structure in one embodiment of this application showing micropillars disposed on a sorting chip;

[0062] Figure 15 : A schematic diagram of a wave-shaped structure of the sorting chip in one embodiment of this application;

[0063] Figure 16 : A schematic diagram of the structure of an apparatus for sorting and enrichment in another embodiment of this application.

[0064] Explanation of reference numerals in the attached figures:

[0065] 10a. Upper shell; 11a. Side wall of upper shell;

[0066] 20a, Lower shell; 21a, Side wall of lower shell;

[0067] 30a, Sorting chip; 31a, First sorting chip; 32a, Second sorting chip; 33a, Third sorting chip; 34a, Screening hole; 35a, Chip sidewall;

[0068] 41a, First cavity; 42a, Second cavity; 43a, Third cavity; 44a, First outlet; 45a, Second outlet; 46a, Third outlet; 47a, First inlet; 48a, Third inlet;

[0069] 50a, Opening; 60a, Microcolumn; 70a, Cell Cluster.

[0070] 10b. Upper shell;

[0071] 20b. Lower housing;

[0072] 31b, First sorting chip; 32b, Second sorting chip; 34b, Screening hole;

[0073] 41b, First cavity; 42b, Second cavity; 43b, Third cavity; 44b, First outlet; 45b, Second outlet; 46b, Third outlet; 48b, Third inlet;

[0074] 70b, Cell clusters. Detailed Implementation

[0075] The following embodiments of this application are only used to illustrate specific implementation methods of this application, and these embodiments should not be construed as limitations on this application. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of this application shall be considered equivalent substitutions and fall within the protection scope of this application.

[0076] This embodiment provides an apparatus for sorting and enriching, such as... Figures 1-11 As shown, it includes:

[0077] case;

[0078] N sorting chips, each with a screening hole 34a, are arranged sequentially to divide the internal space of the housing into N+1 cavities, where N≥1. Figure 1 As shown, in this embodiment, the N sorting chips are three sorting chips arranged from top to bottom—the first sorting chip 31a, the second sorting chip 32a, and the third sorting chip 33a; wherein,

[0079] 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);

[0080] A second cavity 42a is formed between the other side of the housing (the inner side of the lower housing 20a) and the adjacent sorting chip (such as the third sorting chip 33a);

[0081] The cavity wall of the first cavity 41a has a first outlet 44a and a first inlet 47a that connect the first cavity 41a with the outside.

[0082] A second outlet 45a is formed on the cavity wall of the second cavity 42a, connecting the second cavity 42a with the outside.

[0083] There are no particular limitations on the composition of the housing in this application, as long as it can form a sealed internal space. For example, it can be a separate closed housing with each sorting chip arranged sequentially inside it, or it can be composed of the upper housing 10a, the lower housing 20a and the chip sidewalls 35a of the N sorting chips 30a arranged between them, as in this embodiment.

[0084] Regarding the specific implementation of dividing the internal space of the housing into N+1 cavities by sequentially arranged N sorting chips, this application does not impose specific restrictions. For example, if N sorting chips are sequentially arranged inside a closed housing, the N sorting chips will divide the internal space of the housing into N+1 cavities. Alternatively, as in this embodiment, the upper housing 10a, sorting chip 30a, and / or lower housing 20a are recessed, and the N+1 cavities are formed by stacking the upper housing 10a, lower housing 20a, and sorting chip 30a (see reference). Figures 1-9 ).

[0085] Regarding the specific implementation of each outlet (such as the first outlet 44a, the second outlet 45a, and the third outlet 46a hereinafter) and / or inlet (the first inlet 47a and the second and third inlets 48a hereinafter), this application does not impose specific restrictions. For example, a channel connecting each cavity to the outside can be directly opened on the housing to obtain the corresponding inlet and / or outlet. Alternatively, as in this embodiment, an opening 50a can be provided on the side wall of the upper housing 10a, the sorting chip 30a, and / or the lower housing 20a, and the corresponding inlet and / or outlet can be obtained by stacking the upper housing 10a, the lower housing 20a, and the sorting chip 30a (see reference). Figures 1-9 ).

[0086] The number and density of each screening hole 34a can be specifically set by those skilled in the art as needed, and will not be elaborated further in this application. Specifically, multiple screening holes 34a can be arranged in a certain density array.

[0087] Regarding the shape of the screening wells, this application does not impose specific limitations, as long as they can be applied to the screening of the corresponding cell clusters. For example, the cross-sectional shape of the screening wells can be circular or polygonal. Figure 10 As shown, the cross-sectional shape of its 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 various shapes, such as... Figure 11 In the sorting chip, the screening holes are arranged in a combination of rectangles and circles, that is, in at least one (such as one, some or all) of the sorting chips, at least two of the screening holes have different shapes and / or apertures.

[0088] Regarding the pore size of each screening well 34a, those skilled in the art can set it specifically as needed. Specifically, in this embodiment, the pore size of the screening well is as small as 8μm, that is, the pore size of the screening well 34a in this application is 8μm or larger, specifically such as 9μm, 10μm, 12μm, 15μm, 18μm, 20μm, 25μm, 30μm, 35μm, 50μm, 75μm, 100μm, 200μm, or 500μm or larger. Further, when the pore size of the screening well 34a is less than or equal to 40μm, the sorting chip is suitable for sorting and / or free cell clusters present in samples rich in mass fluid. It should be noted that, in this application, the pore size refers to the diameter of the largest circle that the cross-section of the screening well can accommodate.

[0089] The housing (such as upper housing 10a, lower housing 20a, etc.) and / or the sorting chip can be fabricated using existing and mature microfluidic chip processing technologies such as etching and injection molding. Then, they are bonded and packaged to form the complete sorting and enrichment device of this application.

[0090] Regarding the materials of the housing (such as upper housing 10a, lower housing 20a, etc.) and / or sorting chips in this application, they can be metallic materials, inorganic materials and / or polymeric materials, preferably inorganic materials (such as glass, silicon wafers, ceramics) and / or polymeric materials (such as polymeric materials).

[0091] When using the device provided in this embodiment, a fluid sample is supplied through the first inlet 47a (here, a fluid sample containing cell clusters is taken as an example; the principle is the same for other fluid samples). Applying positive pressure to the first inlet 47a, negative pressure to the first outlet 44a, and / or negative pressure to the second outlet 45a provides the power 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 small pore sizes in the sorting holes 34a of the sorting chip, they pass through the sorting holes 34a and enter the lower cavity; Movement direction 2: For cells and / or cell clusters with large pore sizes in the sorting holes 34a of the sorting chip, they cannot pass through the sorting holes 34a and can only flow towards the outlet corresponding to their cavity. Therefore, as... Figure 1 As shown, a fluid sample is supplied through the first inlet 47a. Through the aforementioned "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 of the corresponding size range (larger size). Through the aforementioned "movement direction 1", cells and / or cell clusters smaller than the screening holes 34a of the first sorting chip 31a enter the lower cavity through the screening holes 34a of the first sorting chip 31a, thereby further screening and enriching cells and / or cell clusters of the corresponding size range (smaller size).

[0092] It should be noted that, in this application, "cell cluster" refers to a cluster of cells consisting of two or more cells bound together by covalent and / or non-covalent interactions and moving in a fluid as a whole.

[0093] Preferably, when N≥2 (i.e., when there are two or more sorting chips, such as 3, 4, 5, 6, 7, 8, 9 or more, taking the example of three sorting chips in this embodiment), a third cavity 43a is formed between the sorting chips, and a third outlet 46a is formed on the cavity wall of each third cavity 43a to connect the third cavity 43a with the outside. From one side of the housing (upper housing 10a) to the other side of the housing (lower housing 20a), the aperture of the screening hole of each sorting chip gradually decreases. At this time, negative pressure can also be provided through the third outlet 46a to provide power for the flow of fluid samples. The fluid sample flowing downwards from the first cavity 41a into the third cavity 43a (the cavity between the first sorting chip 31a and the second sorting chip 32a) also has two flow directions. In "movement direction 2," cells and / or cell clusters larger than the pore size of the second sorting chip 32a flow into the corresponding third outlet 46a of this cavity, thereby screening and enriching cells and / or cell clusters of the corresponding size range. In "movement direction 1," cells and / or cell clusters smaller than the pore size of the second sorting chip 32a enter the lower cavity through the screening holes 34a of the second sorting chip 32a, thereby screening and enriching cells and / or cell clusters of the corresponding size range. Based on the same principle, from top to bottom, each sorting chip achieves step-by-step screening and enrichment of cells and / or cell clusters of different sizes.

[0094] Preferably, two or more (two in this embodiment) first inlets 47a are formed on the wall of the first cavity 41a; and / or, at least one (such as one, some or all) of the third cavity has a third inlet 48a on its wall connecting the third cavity 43a to the outside. Generally, the screened and enriched cells and / or cell clusters need to be suspended in a corresponding fluid (such as a buffer solution and other components contained in the buffer solution) 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 additional first inlets 47a and / or third inlets 48a, and even positive pressure can be provided by the fluid. The introduced fluid can, on the one hand, drive the flow of fluid samples in the corresponding cavity, reduce / prevent cells and / or cell clusters from clogging the screening pores, and on the other hand, directly collect the 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 purposes.

[0095] Of course, a corresponding second inlet can also be provided on the second cavity 42a to facilitate the above-mentioned effects 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.

[0096] In one embodiment of the above examples, such as Figures 12-14 As shown, the device further includes micropillars 60a (also referred to as "protrusions"), at least one micropillar 60a is provided in at least one (e.g., one, part or all) of the first cavity 41a, the second cavity 42a and the third cavity 43a, more preferably, the micropillars 60a are provided in all of the above cavities.

[0097] For example, the micropillars 60a can be disposed on the inner surface of the housing, so that the micropillars 60a extend into the first cavity 41a, the second cavity 42a, and / or the third cavity 43a. Figure 12 The diagram shows a schematic of a micropillar 60a disposed on the upper housing 10a. The micropillar 60a is disposed on the inner side of the upper housing 10a, specifically on the inner side of the upper housing sidewall 11a and on the inner side of the upper housing 10a facing the adjacent sorting chip (first sorting chip 31a). For example... Figure 13 The diagram shows a schematic of a micropillar 60a disposed on the lower housing 20a. The micropillar 60a is disposed on the inner side of the lower housing 20a, specifically on the inner side wall 21a of the lower housing 20a and on the inner side of the lower housing 20a facing the adjacent sorting chip (third sorting chip 33a). For example, as shown... Figure 14 The diagram shows a micropillar 60a disposed on the sorting chip (specifically the first sorting chip 31a), which is disposed on the inner side of the chip sidewall 35a.

[0098] The micropillar 60a can also be configured to face the sorting chip towards one side of the housing (e.g., towards the upper housing 10a) and / or towards the other side of the housing (e.g., towards the lower housing 20a), such as... Figure 14 The diagram shows a schematic of micropillars 60a disposed on a sorting chip (specifically, the first sorting chip 31a). The micropillars 60a can be disposed on its upper side, or, if necessary, on its lower side. That is, the micropillars 60a can be disposed in the area of ​​the sorting chip used to set the screening holes 34a (i.e., the screening area described below).

[0099] Regarding the number and density of each micropillar 60a, those skilled in the art can make specific settings as needed, which will not be elaborated in this application. Specifically, multiple micropillars 60a can be arrayed in at least one (e.g., one, part or all) of the first cavity 41a, the second cavity 42a, and the third cavity 43a.

[0100] Regarding the cross-sectional shape of the micropillar 60a, it can be set to a circle, ellipse, and / or polygon, etc., as needed. Figures 12-14 Examples of micropillar 60a with hexagonal and trapezoidal cross sections are given in the example.

[0101] By setting the aforementioned micropillars 60a in the cavity, the original flow direction of the fluid in the cavity can be changed, increasing the chance of cells and / or cell clusters 70a in the fluid sample coming into contact with the screening holes 34a of the sorting chip 30a. This allows cell clusters smaller than the screening holes 34a to flow through the screening holes 34a to the next cavity as much as possible, rather than flowing directly to the outlet corresponding to the cavity, thereby enhancing the screening and enrichment function of the device of this application.

[0102] In one embodiment of the above embodiments, 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 in the region where the screening hole 34a is provided; at least one (e.g., one, part, or all) of at least one surface of the screening area of ​​the sorting chip is a non-planar structure, preferably, at least one surface of the screening area of ​​the sorting chip facing the first cavity is a non-planar structure, and more preferably, both surfaces of the screening area of ​​the sorting chip are non-planar structures. Figure 15 An example is given where both surfaces of the filtering area are curved (specifically, wavy).

[0103] By making the surface of the screening area facing the first cavity non-planar, the original flow direction of the fluid in the cavity can be changed, increasing the chance of cell clusters 70a contacting the screening holes 34a of the sorting chip 30a. This allows cell clusters smaller than the screening holes 34a to flow through the screening holes 34a to the next cavity as much as possible, rather than flowing directly to the outlet corresponding to the cavity, thereby enhancing the screening and enrichment function of the device of this application.

[0104] In another embodiment of this application, another apparatus for sorting and enrichment is provided, such as... Figure 16 As shown, it includes:

[0105] case;

[0106] N sorting chips, each with a screening hole 34b, are arranged sequentially to divide the internal space of the housing into N+1 cavities, where N≥2. Figure 16 As shown, in this embodiment, the N sorting chips are two sorting chips arranged from top to bottom—the first sorting chip 31b and the second sorting chip 32b; wherein,

[0107] 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 (i.e., the first sorting chip 31b).

[0108] A second cavity 42b is formed between the other side of the housing (the inner side of the lower housing 20b) and the adjacent sorting chip (such as the second sorting chip 32b);

[0109] 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).

[0110] A first outlet 44b is formed on the cavity wall of the first cavity 41b, connecting the first cavity 41b with the outside.

[0111] A second outlet 45b is formed on the cavity wall of the second cavity 42b, connecting the second cavity 42b with the outside.

[0112] Each of the third cavities 43b has a third outlet 46b on its cavity wall that connects the third cavity 43b to the outside, and at least one (such as one, some or all) of the third cavities 43b has one or more third inlets 48b on its cavity wall that connect the third cavity 43b to the outside.

[0113] In this embodiment, the composition and structure of the housing; the specific implementation of dividing the internal space of the housing into N+1 cavities by the N sequentially arranged sorting chips; the specific implementation 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 materials of the housing and / or sorting chips; the setting position, size, density and shape of the micropillars; the surface form of the screening area of ​​the sorting chip, etc., are all the same as in the previous embodiment, and those skilled in the art will know that they can also bring the same effect, so they will not be repeated here.

[0114] When using the device provided in this embodiment, a fluid sample is supplied through the third inlet 48b (here, a fluid sample containing cell clusters is taken as an example; the principle is the same for other fluid samples). Applying positive pressure to the third inlet 48b, negative pressure to the first outlet 44b, negative pressure to the second outlet 45b, and / or negative pressure to the third outlet 46b provides the power 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 small pore sizes compared to the sorting chip's screening holes 34b, they pass through the screening holes 34b and enter the adjacent cavity; Movement direction 2, for cells and / or cell clusters with large pore sizes compared to the sorting chip's screening holes 34b, they cannot pass through the sorting chip's screening holes 34b and can only flow towards the outlet (third outlet 46b) of their respective cavity. Therefore, if... Figure 16As shown, a fluid sample is supplied through the third inlet 48b. Within the third cavity 43b, cells and / or cell clusters larger than the screening holes 34b of the sorting chips (such as the first sorting chip 31b and the second sorting chip 32b) on both sides of the third cavity 43b flow into the third outlet 46b via the aforementioned "movement direction 2," thereby screening and enriching cells and / or cell clusters of the corresponding size range (larger size). Through the aforementioned "movement direction 1," cells and / or cell clusters smaller than the screening holes 34b of the sorting chips (such as the first sorting chip 31b and the second sorting chip 32b) on both sides enter the cavities (such as the first cavity 41b or the second cavity 42b) through the screening holes 34b of the first sorting chip 31b or the second sorting chip 32b, thereby further screening and enriching cells and / or cell clusters of the corresponding size range (smaller size).

[0115] When the number of sorting chips on either side of the third inlet 48b used for introducing fluid samples is greater than two, the aperture of the screening holes 34b between the sorting chips can be gradually reduced along the direction away from the third inlet 48b used 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.

[0116] Preferably, at least one of the third cavities 43b has two or more third inlets 48b forming on its cavity wall, connecting the third cavity to the outside. Generally, after screening and enrichment, cells and / or cell clusters need to be suspended in a corresponding fluid (such as a buffer solution and other fluids 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, while the additional third inlets 48b are used to introduce the corresponding fluid, and may even provide positive pressure through this fluid. The introduced fluid can, on the one hand, drive the flow of the fluid sample within the corresponding cavity, reducing / preventing cell and / or cell cluster blockage of the screening pores; on the other hand, it can directly collect the 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 purposes. Of course, in addition to providing two or more third inlets 48b on one third cavity 43b, other cavities (such as the first cavity 41b, the second cavity 42b and / or other third cavities 43b) are provided with corresponding inlets (such as a first inlet on the first cavity; a second inlet on the second cavity; and / or, a third inlet is also provided in at least one (such as one, part or all) of other third cavities), so as to introduce the corresponding fluid through the corresponding inlet, so as to facilitate the above-mentioned effects 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.

[0117] Preferably, N=2, and the pore sizes of the screening holes 34b between the sorting chips (first sorting chip 31b and second sorting chip 32b) on both sides of the third cavity 43b are the same. In the fluid sample introduced through the third inlet 48b, cells and / or cell clusters smaller than the pore size of the screening holes 34b enter the first and second cavities through the screening holes 34b of the sorting chips, and flow out and are collected through the corresponding first outlet 44b and second outlet 45b; cells and / or cell clusters larger than the pore size of the screening holes 34b are retained in the third cavity by the sorting chip and flow out and are collected through the corresponding third outlet 46b. 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 this device for cell and / or cell cluster sorting, the cells and / or cell clusters trapped by the sorting chip can easily flow directly out of the device with the solvent through the third outlet, which is less likely to clog the screening holes 34b of the sorting chip, thus improving the sorting and / or enrichment efficiency of the device. In addition, the device has two sorting chips with the same pore diameter of screening holes 34b, which also helps to improve the sorting and enrichment efficiency of the device, making the device suitable for processing large volume samples.

[0118] Any of the above devices can be used to sort and enrich the following fluid samples:

[0119] (1) Cell clusters in peripheral blood samples;

[0120] (2) Cell clusters in pleural effusion, ascites, lymph, urine, or cerebrospinal fluid;

[0121] (3) Cell clusters formed after enzymatic digestion of solid tissue; and / or,

[0122] (4) Cell clusters formed by the re-aggregation of single cells after digestion of solid tissue into single cells.

[0123] In addition, it can also be used to sort and enrich mixtures of particles of different sizes containing other biomolecules, such as (5) liposomes, water-in-oil microdroplets or oil-in-water microdroplets.

[0124] That is, in addition to providing any of the above-mentioned devices, this application also provides the application of the above-mentioned devices in sorting and enriching the above-mentioned fluid samples.

[0125] 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 of specific structures, connection relationships, etc.; in addition, the orientation or positional relationship indicated by "longitudinal," "lateral," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," etc., is based on the orientation or positional relationship shown in the accompanying drawings, and is only for the convenience of describing this application and simplifying the description, and is not intended to indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, the above terms should not be construed as limitations on this application.

[0126] Although the embodiments of this application have been described above, this application is not limited to the specific embodiments and application fields described above. The specific embodiments described above are merely illustrative and instructive, and not restrictive. Those skilled in the art, based on the guidance of this specification and without departing from the scope of protection of the claims of this application, can make many other forms, all of which are within the scope of protection claimed in this application.

Claims

1. An apparatus for sorting and enriching, wherein, The device comprises: a housing; N sorting chips, each of which is provided with a screening hole, and N sorting chips arranged in sequence divide the internal space of the housing into N+1 cavities, N≥1; wherein, 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 with the outside is formed on the cavity wall of the second cavity.

2. The device of claim 1, wherein, N≥2； a third cavity is formed between any two 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 of claim 2, wherein, at least one third inlet connecting the third cavity with the outside is formed on the cavity wall of at least one third cavity.

4. The device of claim 1, wherein, more than two first inlets are formed on the cavity wall of the first cavity.

5. The device of claim 2, wherein, the aperture of the screening hole of each sorting chip gradually decreases from one side of the housing to the other side of the housing; and / or, the aperture of the screening hole is greater than or equal to 8 μm.

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

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

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

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

10. The device of claim 1, wherein, the region where the sorting chip is provided with the screening hole forms a screening area; at least one surface of the screening area of at least one sorting chip is partially or entirely non-planar.

11. The device of claim 10, wherein, at least one surface of the screening area of at least one sorting chip facing the first cavity is partially or entirely non-planar.

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

13. An apparatus for sorting and enriching, wherein, The device comprises: a housing; N sorting chips, each of which is provided with a screening hole, and N sorting chips arranged in sequence divide the internal space of the housing into N+1 cavities, N≥2; wherein, 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 is formed on a cavity wall of the first cavity for connecting the first cavity with the outside world. A second outlet is formed on a cavity wall of the second cavity for connecting the second cavity with the outside world. A third outlet is formed on a cavity wall of each of the third cavities for connecting the third cavities with the outside world, and at least one of the third cavities has more than one third inlet formed on a cavity wall thereof for connecting the third cavity with the outside world.

14. The apparatus of claim 13, wherein, N = 2; and / or, At least one of the third cavities has more than two third inlets formed on a cavity wall thereof for connecting the third cavity with the outside world.

15. The apparatus of claim 14, wherein, N = 2; and the apertures of the screening holes of the two sorting chips are identical.

16. The apparatus of claim 13, wherein, The apertures of the screening holes are equal to or greater than 8 μιη.

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

18. The apparatus of claim 13, wherein, The apparatus further comprises a micro-pillar, and at least one micro-pillar is disposed in at least one of the first cavity, the second cavity, and the third cavities.

19. The apparatus of claim 18, wherein, The micro-pillar is disposed on the housing; and / or, The micro-pillar is disposed on a surface of the sorting chip.

20. The apparatus of claim 19, wherein, The cross-section of the micro-pillar is circular, elliptical, and / or polygonal.

21. The apparatus of claim 13, wherein, The sorting chip forms a screening area in the region of the screening holes; At least one surface of the screening area of at least one of the sorting chips is partially or entirely non-planar.

22. The apparatus of claim 21, wherein, At least one surface of the screening area of at least one of the sorting chips facing the third inlet is partially or entirely non-planar.

23. The apparatus of claim 21, wherein, The non-planar surface is a curved surface.

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