Cell screening chip, cell screening system and method thereof

Abstract

The present disclosure provides a cell screening chip, a cell screening system and a method thereof, which relate to the field of biological detection technology and are used to solve the technical problems of long separation time and low separation efficiency of target cells. The cell screening chip has a plurality of screening units formed in the partition between the inlet groove and the outlet groove of the chip body, each screening unit includes a containing cavity and a screening channel, and the inlet groove, the screening unit and the outlet groove are connected in sequence; in the second surface of the cover covering the first surface of the partition, the minimum value of the first distance between the area corresponding to the screening channel and the first surface of the partition is less than the radius of the target cell, and the maximum value is less than the diameter of the target cell, and the maximum value of the second distance between other areas and the first surface of the partition is less than the diameter of the target cell, and the sample solution is diverted between the cover and the partition to shorten the separation time and improve the separation efficiency.

Description

Technical Field

[0001] The present disclosure relates to the field of biological detection technology, and in particular to a cell screening chip, a cell screening system and a method thereof. Background Art

[0002] In the field of biological detection, it is often necessary to separate target cells for further observation or testing of target cells, for example, separation and detection of circulating tumor cells. Circulating tumor cells (CTCs) are tumor cells that originate from primary tumors and enter the blood circulation system. These cells can develop into metastatic lesions through the blood circulation system and then multiply to form secondary tumors. Therefore, timely separation and detection of these cells is of great significance for monitoring tumor treatment and recurrence.

[0003] For the screening of target cells, the difference in physical properties between target cells and other cells can be used to separate target cells. For example, a microfilter with a channel size smaller than the diameter of the target cells can be designed to capture larger target cells and allow other smaller cells to flow out with the sample solution, thereby separating the target cells. However, when this method is used to separate target cells from the sample solution, it takes a long time to separate the target cells from the sample solution and the separation efficiency is low. Summary of the invention

[0004] In view of the above problems, the embodiments of the present disclosure provide a cell screening chip, a cell screening system and a method thereof, which are used to shorten the separation time of target cells and improve the separation efficiency of target cells.

[0005] In order to achieve the above objectives, the embodiments of the present disclosure provide the following technical solutions:

[0006] In a first aspect, an embodiment of the present disclosure provides a cell screening chip, comprising a chip body and a cover, wherein the chip body comprises a liquid inlet groove and a liquid outlet groove, and a partition formed between the liquid inlet groove and the liquid outlet groove, wherein a screening array is formed in the partition, and the screening array comprises a plurality of screening units arranged along the extension direction of the liquid inlet groove; each of the screening units comprises a accommodating cavity and a screening channel, wherein the inlet end of the accommodating cavity is connected to the liquid inlet groove, the outlet end of the accommodating cavity is connected to the inlet end of the screening channel, and the outlet end of the screening channel is connected to the liquid outlet groove; the second surface of the cover is arranged on the first surface of the partition, and in the second surface of the cover, the minimum value of the first distance between the area corresponding to the screening channel and the first surface of the partition is less than the radius of the target cell, and the maximum value of the first distance is less than the diameter of the target cell; the maximum value of the second distance between other areas in the second surface of the cover and the first surface of the partition is less than the diameter of the target cell.

[0007] In the cell screening chip provided by the embodiments of the present disclosure, the chip body includes a liquid inlet groove and a liquid outlet groove, a screening array is formed in the partition between the liquid inlet groove and the liquid outlet groove, the screening array includes a plurality of screening units, each screening unit includes a accommodating cavity and a screening channel, the liquid inlet groove, the accommodating cavity, the screening channel and the liquid outlet groove are connected in sequence, the sample solution containing target cells flows into the accommodating cavity and the screening channel from the liquid inlet groove, and flows out from the liquid outlet groove; the separation of target cells and non-target cells is achieved in the screening unit; at the same time, the first surface of the partition is covered with the second surface of the cover, a gap is formed between the cover and the partition, and the area corresponding to the screening channel in the second surface of the cover is aligned with the first surface of the partition The minimum value of the first distance between the two surfaces is smaller than the radius of the target cell, and the maximum value of the first distance is smaller than the diameter of the target cell, which can prevent the target cell from leaking out from the connection between the gap and the screening channel; and the maximum value of the second distance between other areas in the second surface of the cover and the first surface of the partition is smaller than the diameter of the target cell, which can prevent the target cell from leaking out from the gap, thereby ensuring that the cell screening chip can work normally. Compared with the prior art in which the cover is in contact with the partition, the sample solution can be diverted between the cover and the partition in the embodiment of the present disclosure, which increases the flow rate of the sample solution, thereby shortening the separation time of the target cells and improving the separation efficiency of the target cells.

[0008] In a second aspect, an embodiment of the present disclosure further provides a cell screening system, comprising an injection pump, a waste liquid collection device, and the cell screening chip as described above, wherein the injection port of the cell screening chip is connected to the injection pump, and the outlet of the cell screening chip is connected to the waste liquid collection device.

[0009] The cell screening system provided by the embodiments of the present disclosure includes the above-mentioned cell screening chip, and thus also has the advantages of short separation time and high separation efficiency of target cells. The specific effects are referred to above and will not be repeated here.

[0010] In a third aspect, an embodiment of the present disclosure also provides a cell screening method, comprising: injecting a surface treatment liquid and a buffer solution into a cell screening chip in sequence for pretreatment; injecting a sample solution containing target cells into the cell screening chip, wherein the cell screening chip captures the target cells; injecting a buffer solution, a fixative solution, a buffer solution, a staining solution and a buffer solution into the cell screening chip in sequence to fix and stain the target cells; an image acquisition device acquires an image of the cell screening chip and transmits the image to a data processing device, wherein the data processing device identifies the target cells.

[0011] In the embodiment of the present disclosure, the cell screening chip is pre-treated and then injected with a sample solution, and the target cells in the sample solution are captured by the cell screening chip. Since the cell screening method is a method corresponding to the above-mentioned cell screening system, the separation time of the target cells can be shortened and the separation efficiency of the target cells can be improved, which will not be described in detail here. At the same time, the target cells are fixed with a fixing solution, and the target cells are stained and identified with a staining solution to distinguish them from non-target cells, which is convenient for the identification of the target cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is a schematic diagram of the first structure of the cell screening chip in the embodiment of the present disclosure;

[0013] Figure 2 It is a partial structural schematic diagram of a cell screening chip in an embodiment of the present disclosure;

[0014] Figure 3 A schematic diagram of the structure of a chip body in an embodiment of the present disclosure;

[0015] Figure 4 for Figure 3 A partial enlarged view of the middle A;

[0016] Figure 5 Another structural diagram of the chip body in the embodiment of the present disclosure is shown in FIG.

[0017] Figure 6 is a second structural schematic diagram of a cell screening chip in an embodiment of the present disclosure;

[0018] Figure 7 Schematic diagram of the third structure of the cell screening chip in the embodiment of the present disclosure;

[0019] Figure 8Schematic diagram of the fourth structure of the cell screening chip in the embodiment of the present disclosure;

[0020] Fig. 9 is a schematic diagram of the flow of the sample solution in the chip body in the embodiment of the present disclosure;

[0021] Fig.10 is a schematic structural diagram of a buffer tank in an embodiment of the present disclosure;

[0022] Fig.11 for Fig.10 A partial enlarged view of point B in the middle;

[0023] Fig.12 A schematic diagram of the structure of a multi-screening channel in an embodiment of the present disclosure;

[0024] Fig.13 for Fig.12 A partial enlarged view of point C in the middle;

[0025] Fig.14 Another structural schematic diagram of a multi-screening channel in an embodiment of the present disclosure;

[0026] Fig.15 for Fig.14 A partial enlarged view of point D in the middle;

[0027] Fig.16 is a schematic diagram of the distribution of accommodating cavities of different widths in an embodiment of the present disclosure;

[0028] Fig.17 It is a schematic diagram of a first structure of a liquid inlet groove in an embodiment of the present disclosure;

[0029] Fig.18 for Fig.17 The first partial enlarged view at E in the middle;

[0030] Fig.19 for Fig.17 The second partial enlarged view at E in the middle;

[0031] Fig. 20 for Fig.17 The third partial enlarged view at E in the middle;

[0032] Fig.21 is a schematic diagram of a second structure of the liquid inlet groove in the embodiment of the present disclosure;

[0033] Fig. 22 Schematic diagram of the third structure of the liquid inlet groove in the embodiment of the present disclosure;

[0034] Fig.23 is a fourth structural schematic diagram of the liquid inlet groove in the embodiment of the present disclosure;

[0035] Fig.24 It is a schematic diagram of the first structure of the dual injection ports in the embodiment of the present disclosure;

[0036] Fig.25 It is a schematic diagram of the second structure of the dual injection port in the embodiment of the present disclosure;

[0037] Fig.26 is a structural schematic diagram of a liquid inlet connecting pipeline in an embodiment of the present disclosure;

[0038] Fig. 27 is a schematic diagram of the structure of a cell screening system in an embodiment of the present disclosure;

[0039] Fig.28 Flow chart of the cell screening method in an embodiment of the present disclosure.

[0040] Description of reference numerals:

[0041] 10. Cell screening chip; 100. Chip body;

[0042] 110, injection port; 111, first injection port;

[0043] 112, a second sample inlet; 120, a liquid inlet groove;

[0044] 121. straight channel; 122. first arc channel;

[0045] 123. arc-shaped guide plate; 124. second arc-shaped channel;

[0046] 130. Screening array; 131. Accommodating cavity;

[0047] 132. Screening channel; 133. Buffer tank;

[0048] 140, liquid outlet groove; 150, sample outlet;

[0049] 160. Liquid inlet connecting pipe; 161. Liquid inlet diversion pipe;

[0050] 162. First-stage liquid inlet diversion pipeline; 163. Second-stage liquid inlet diversion pipeline;

[0051] 170. Liquid outlet connecting pipe; 171. Liquid outlet diversion pipe;

[0052] 180. Cover; 181. Raised portion;

[0053] 190, separator;

[0054] 191. separator block; 192. extension portion;

[0055] 20. Sample injection pump; 21. Surface treatment liquid pump;

[0056] 22. Sample solution pump; 23. Buffer solution pump;

[0057] 24. Fixative liquid pump; 25. Dyeing liquid pump;

[0058] 30. Directional valve; 40. Light source;

[0059] 50. Image acquisition device; 60. Data processing device;

[0060] 70. Waste liquid collection device; 80. Carrier;

[0061] L1, the width of the accommodating cavity; L2, the width of the screening channel;

[0062] K, the second surface of the cover corresponds to the area of ​​the screening channel;

[0063] a, angle; d, minimum value of the first distance;

[0064] h. The maximum value among the second distances. DETAILED DESCRIPTION

[0065] In the related art, a cell screening chip includes a chip body and a cover, which are sealed together, and a cavity for screening target cells is provided between the chip body and the cover. However, the cell screening chip has technical problems such as long separation time and low separation efficiency of target cells.

[0066] In view of the above technical problems, the cell screening chip in the embodiment of the present disclosure includes a chip body and a cover, and the partition between the liquid inlet groove and the liquid outlet groove of the chip body forms a screening array, and the screening array includes a plurality of screening units, each screening unit includes a receiving cavity and a screening channel, and the liquid inlet groove, the receiving cavity, the screening channel and the liquid outlet groove are connected in sequence. The second surface of the cover is covered on the first surface of the partition, and the minimum value of the first distance between the area corresponding to the screening channel and the first surface of the partition in the second surface of the cover is less than the radius of the target cell, the maximum value of the first distance is less than the diameter of the target cell, and the maximum value of the second distance between other areas and the first surface of the partition is less than the diameter of the target cell, so that the sample solution can be diverted between the cover and the partition, and the target cells are prevented from flowing out from between the cover and the partition, thereby increasing the flow rate of the sample solution, thereby shortening the separation time of the target cells, and improving the separation efficiency of the target cells.

[0067] In order to make the above-mentioned purposes, features and advantages of the embodiments of the present disclosure more obvious and understandable, the technical solutions in the embodiments of the present disclosure will be clearly and completely described below in combination with the drawings in the embodiments of the present disclosure. Obviously, the described embodiments are only part of the embodiments of the present disclosure, not all of the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present disclosure.

[0068] Embodiment 1

[0069] In order to separate target cells in a sample solution for subsequent observation or testing, an embodiment of the present disclosure provides a cell screening chip. The cell screening chip may be a microfluidic chip, and its material may be transparent PMMA, PC, PS or glass, so as to facilitate subsequent identification of target cells.

[0070] Reference Figure 1 and Figure 2 The cell screening chip in the embodiment of the present disclosure includes a chip body 100 and a cover 180. The cover 180 is covered on the chip body to seal the chip body 100 and prevent the sample solution from flowing out of the cell screening chip. The cover 180 can be located above the chip body 100 or below the chip body 100. Figure 1 and Figure 2 As shown, the cover 180 covers the chip body 100, and the bottom surface of the cover 180 is in contact with the upper surface of the chip body 100. This arrangement can prevent the target cells from overflowing from the gap between the two surfaces into the target cell screening area in the chip body 100, thereby improving the interception rate of the target cells and further improving the detection accuracy of the cell screening chip.

[0071] The cover 180 may be a flat plate structure to facilitate processing and molding of the cover 180. The size of the bottom surface of the cover 180 may be consistent with the size of the top surface of the chip body 100, such as Figure 1 and Figure 2 As shown, the cover 180 covers the entire upper surface of the chip body 100. The size of the bottom surface of the cover 180 can also be smaller than the size of the upper surface of the chip body 100. The cover 180 covers part of the upper surface of the chip body 100, that is, covers the functional area of ​​the chip body 100, so as to reduce the volume of the cover 180.

[0072] The cover 180 may be injection molded, and the cover 180 and the chip body 100 are bonded, for example, by thermal bonding, adhesive bonding, or ultrasonic bonding. The cover 180 may also be a film layer formed on the chip body 100 by a film lamination process. The cover 180 may also be integrally formed with the chip body 100, for example, the cover 180 and the chip body 100 are integrally formed by 3D printing.

[0073] The chip body 100 may also be formed by injection molding. When the chip body 100 is formed by injection molding, a mold of the chip body 100 needs to be made first, and the mold may be formed by electroforming, machining or etching. The chip body 100 may also be manufactured by other micro-manufacturing technologies such as laser etching and photolithography.

[0074] Reference Figures 1 to 3 The chip body 100 is provided with an injection port 110, a liquid inlet groove 120, a screening array 130, a liquid outlet groove 140 and a sample outlet 150. One end of the liquid inlet groove 120 is connected to the injection port 110, and the other end is blocked. For example, along the flow direction of the liquid inlet groove 120, the head end of the liquid inlet groove 120 is connected to the injection port 110, and the end of the liquid inlet groove 120 is blocked. The liquid outlet groove 140 is provided on the side of the liquid inlet groove 120, and the screening array 130 is provided between the liquid inlet groove 120 and the liquid outlet groove 140, and is connected by the screening array 130.

[0075] Specifically, the liquid inlet groove 120 and the liquid outlet groove 140 can be arranged in parallel. A screening array 130 is arranged between the side walls of the liquid inlet groove 120 and the liquid outlet groove 140 close to each other, and the screening array 130 is connected. The side walls of the liquid inlet groove 120 and the liquid outlet groove 140 close to each other refer to the side wall of the liquid inlet groove 120 close to the liquid outlet groove 140 and the side wall of the liquid outlet groove 140 close to the liquid inlet groove 120.

[0076] One end of the liquid outlet groove 140 is connected to the sample outlet 150, and the other end is blocked. For example, along the flow direction of the liquid outlet groove 140, the head end of the liquid outlet groove 140 is blocked, and the tail end of the liquid outlet groove 140 is connected to the sample outlet 150. The sample solution flows in from the inlet 110, passes through the liquid inlet groove 120, the screening array 130 and the liquid outlet groove 140 in sequence and flows to the sample outlet 150. The screening array 130 is used to intercept and capture target cells.

[0077] In the embodiment of the present disclosure and the following embodiments, the diversion direction of the liquid inlet groove 120 is also the extension direction of the liquid inlet groove 120 , and the diversion direction of the liquid outlet groove 140 is also the extension direction of the liquid outlet groove 140 .

[0078] In the embodiment of the present invention, the liquid inlet groove 120 is a groove body, and the liquid inlet groove 120 has an opening. The bottom surface of the liquid inlet groove 120 refers to the surface opposite to the opening of the liquid inlet groove 120, the side wall of the liquid inlet groove 120 refers to the surface parallel to the extension direction of the liquid inlet groove 120, and the end of the liquid inlet groove 120 refers to the surface starting or ending in the extension direction of the liquid inlet groove 120. It can be understood that the side wall of the liquid inlet groove 120 connects the two ends of the liquid inlet groove 120.

[0079] by Figure 1 Taking the structure shown as an example, the left and right sides of the liquid inlet groove 120 are the ends of the liquid inlet groove 120, the front and rear vertical sides of the liquid inlet groove 120 are the side walls of the liquid inlet groove 120, and the surface of the liquid inlet groove 120 parallel to the upper surface of the chip body 100 is the bottom surface of the liquid inlet groove 120. The structure of the liquid outlet groove 140 can refer to the structure of the liquid inlet groove 120, and will not be repeated here.

[0080] The liquid inlet groove 120 may be provided with a liquid outlet groove 140 on one side or on both sides. That is, at least one of the two sides of the liquid inlet groove 120 is provided with a liquid outlet groove 140, so that a longer screening array 130 can be provided between the liquid inlet groove 120 and the liquid outlet groove 140, thereby improving the capture rate of target cells.

[0081] The inlet groove 120 and the outlet groove 140 may be grooves formed in the chip body 100, and the openings of the inlet groove 120 and the outlet groove 140 face the cover 180. A partition 190 is provided between the inlet groove 120 and the outlet groove 140, and a screening array 130 is formed in the partition 190. For example, Figures 1 to 3 As shown, the partition 190 can be roughly U-shaped, with the open end of the U-shaped partition 190 facing the injection port 110, the bottom end opposite to the open end blocks the liquid inlet groove 120, and the two side walls connected to the two ends of the bottom end respectively form a screening array 130. Figure 3 The right end of the partition 190 shown blocks the liquid inlet groove 120 , and the screening arrays 130 are formed at the upper and lower ends of the partition 190 .

[0082] The partition 190 between the inlet groove 120 and the outlet groove 140 may include a plurality of partition blocks 191, which are arranged along the flow direction of the inlet groove 120, and a screening unit is formed between adjacent partition blocks 191. That is, the screening array 130 includes a plurality of screening units, each of which is not connected to each other and is separated by the partition blocks 191. For example, the screening array 130 includes seven screening units, and the spacing between the screening units is equal.

[0083] like Figure 1 and Figure 2As shown, the upper end surfaces of each partition block 191 are flush to form the first surface of the partition portion 190, and no screening unit is provided in the partition portion 190 located at the right end of the liquid inlet groove 120 to prevent the target cells from entering the liquid inlet groove 120 and accumulating at the end of the liquid inlet groove 120.

[0084] Reference Figure 4 Each screening unit includes a receiving chamber 131 and a screening channel 132 connected to the receiving chamber 131, and the liquid inlet groove 120, the receiving chamber 131, the screening channel 132 and the liquid outlet groove 140 are connected in sequence. That is, the liquid inlet groove 120 is connected to the inlet end of the receiving chamber 131, the outlet end of the receiving chamber 131 is connected to the inlet end of the screening channel 132, and the outlet end of the screening channel 132 is connected to the liquid outlet groove 140.

[0085] Continue to refer to Figure 4 In each screening unit, the width L1 of the accommodating cavity 131 is greater than the diameter of the target cell, and the width L2 of the screening channel 132 is less than the diameter of the target cell. Since the multiple cells in the target cell have different sizes, the diameter of the target cell is usually a range of values. In this embodiment and the following embodiments, the diameter of the target cell refers to the minimum value of the diameter range of the target cell, that is, the diameter value of the smallest cell in the target cell.

[0086] For example, when the target cells are circulating tumor cells, the diameter of the target cells is about 10-20 μm, the width L2 of the screening channel 132 is less than 10 μm, and may be 8 μm, and the width L1 of the accommodating cavity 131 is greater than 10 μm, and may be 20 μm or greater. In this way, the target cells cannot pass through the screening channel 132 and are trapped in the accommodating cavity 131, and non-target cells with a diameter smaller than the screening channel 132 can flow out of the screening channel 132, thereby separating the target cells from the sample solution and trapping them in the accommodating cavity 131. Preferably, a single target cell is captured and trapped in each accommodating cavity 131.

[0087] In each screening unit, the accommodating cavity 131 may be a accommodating groove formed in the partition 190, the opening of the accommodating groove faces the cover 180, and the width of the accommodating cavity 131 refers to the distance between two opposite side walls of the accommodating groove, that is, the distance between two parts of the two partition blocks 191 corresponding to the accommodating cavity 131, such as Figure 4 The length of L1 is shown.

[0088] In each screening unit, the screening channel 132 may be a guide groove or a guide hole formed in the partition 190. When the screening channel 132 is a guide groove, the opening of the guide groove faces the cover 180, and the width of the screening channel 132 refers to the distance between the two opposite side walls of the guide groove, that is, the distance between the two parts of the two partition blocks 191 corresponding to the screening channel 132, such as Figure 4 The length of L2 is shown.

[0089] When the screening channel 132 is a guide hole, the guide hole can be a square hole, a round hole, a semicircular hole, etc. Figure 5 As shown, the guide hole is a rectangular hole, and the guide hole is opened at one end of the partition 190 away from the cover 180. In this way, the height of the screening channel 132 is reduced. When the target cell enters the accommodating cavity 131, the target cell can block or block the entrance end of the screening channel 132, that is, Figure 5 The larger target cells in the screening channel 132 block or block the entrance end of the screening channel 132, reducing the flow in the screening channel 132, or even making the screening channel 132 no longer divert the flow, so that the accommodating chamber 131 reduces or ends the target cell capture function. In addition, it can also prevent the target cells in the accommodating chamber 131 from being squeezed out of the screening channel 132 by the sample solution, thereby improving the interception rate of the accommodating chamber 131.

[0090] Continue to refer to Figures 1 to 3 There is a gap between the partition 190 between the liquid inlet groove 120 and the liquid outlet groove 140 and the cover 180 to divert the sample solution. Specifically, the cover 180 covers the partition 190, and the surface of the cover 180 facing the partition 190 is the second surface of the cover 180. The surface of the partition 190 corresponding to the second surface of the cover 180 is the first surface of the partition 190, that is, the surface of the partition 190 facing the cover 180 is the first surface of the partition 190.

[0091] In the second surface of the cover 180, the minimum value of the first distance between the area corresponding to the screening channel and the first surface of the partition 190 is smaller than the radius of the target cell, and the maximum value of the first distance is smaller than the diameter of the target cell, so that the target cell will not leak out from the connection between the gap and the screening channel 132. The maximum value of the second distance between the other areas in the second surface of the cover 180 and the first surface of the partition 190 is smaller than the diameter of the target cell, so that the target cell will not leak out from the gap, ensuring that the cell screening chip can work normally.

[0092] It can be understood that the cover 180 has a certain area in the area corresponding to the screening channel, and there may be multiple vertical distances between the area and the first surface of the partition 190, and these vertical distances are the first distances. The minimum value in the first distance is the distance between the area closest to the first surface of the partition 190 in the area corresponding to the screening channel and the first surface of the partition 190. By limiting the minimum value in the first distance, it is possible to prevent target cells from leaking out of the connection between the gap and the screening channel 132. The maximum value in the first distance is the distance between the area farthest from the first surface of the partition 190 in the area corresponding to the screening channel and the first surface of the partition 190. By controlling the maximum value in the first distance, it is possible to prevent target cells from leaking out of the gap.

[0093] The second distance refers to the distance between the area of ​​the second surface of the cover 180 that does not correspond to the screening channel and the first surface of the partition 190. The maximum value in the second distance is the distance between the area of ​​the second surface of the cover 180 that is farthest from the first surface of the partition 190 among other areas and the first surface of the partition 190. The maximum value in the second distance is smaller than the diameter of the target cell, so that the target cell will not leak out from the gap.

[0094] It is understandable that the maximum distance of the gap formed between the second surface of the cover 180 and the partition 190 is smaller than the diameter of the target cell, and the minimum distance is smaller than the radius of the target cell, so that the diversion is achieved on the basis of ensuring that the target cell can be retained. In the cell screening chip of the embodiment of the present disclosure, the gap between the screening array 130, the partition 190 and the cover 180 provides more diversion paths for the sample solution, thereby increasing the flow rate of the sample solution in the cell screening chip. Compared with the cell screening chip with only the screening array 130, the embodiment of the present disclosure increases the flow rate of the sample solution and shortens the overall detection time.

[0095] In the above embodiment, the second surface of the cover 180 may be a plane or a curved surface, and the target cells can be intercepted and the solution can be diverted at the gap through the relative position relationship between the cover 180 and the partition 190. Figure 6 , Figure 7 In the specific embodiments, structures of the second surface of the cover 180 having various structures are exemplified, but the present disclosure is not limited thereto.

[0096] In one possible example, referring to Figure 6 , the second surface of the cover 180 is a plane. The lower surface of the cover 180 is a plane, and the first surface of the partition 190 is the upper surface of the partition 190 .

[0097] In the second surface of the cover 180, the area corresponding to the screening channel 132 is Figure 6The distance between the area in K on the second surface of the cover 180 and the first surface of the partition 190 is the first distance, and the minimum value d and the maximum value in the first distance are equal. That is, the first distance is less than or equal to half of the width L2 of the screening channel 132. For example, when the target cells are circulating tumor cells, the width L2 of the screening channel 132 can be 8 μm, Figure 6 The first distance in may be less than or equal to 4 μm.

[0098] In the second surface of the cover 180, the other areas are Figure 6 The maximum value h of the second distance between the area not in K and the first surface of the partition 190 is less than or equal to the width L2 of the screening channel 132. This arrangement prevents the target cells from leaking out from the gap between the cover 180 and the partition 190.

[0099] In another possible example, referring to Figure 7 , the second surface of the cover 180 includes a curved surface, that is, the lower surface of the cover 180 includes a curved surface. The first surface of the partition 190 is the upper surface of the partition 190 .

[0100] In the lower surface of the cover 180, the area corresponding to the screening channel 132 includes a protrusion 181, that is, the protrusion 181 corresponds to the screening channel 132 one by one, and the protrusion 181 is a curved surface. Taking the plane perpendicular to the upper surface of the cover 180 and parallel to the flow direction of the liquid inlet groove 120 as the cross section, the cross-sectional shape of the protrusion 181 can be an arc, a quadrilateral, etc. In the embodiment of the present disclosure, the cross-sectional shape of the protrusion 181 is a semicircle. Figure 7 As shown, the minimum value d of the first distance between the protrusion 181 and the upper surface of the partition 190 is less than or equal to half of the width L2 of the screening channel 132 .

[0101] In the second surface of the cover 180, the distance between the area not corresponding to the screening channel 132 and the first surface of the partition 190 is a second distance, and the maximum value h in the second distance is less than or equal to the width L2 of the screening channel 132. The area of ​​the second surface of the cover 180 not corresponding to the screening channel 132 can be as follows: Figure 7 The plane area shown may also be a concave area away from the partition 190. The target cells are prevented from leaking out from the gap between the cover 180 and the partition 190, and from the connection between the gap and the screening channel 132.

[0102] Reference Figure 8 In order to improve the structural stability of the cover 180 and reduce the possibility of deformation of the cover 180, at least one of the plurality of partition blocks 191 of the partition 190 may be provided with an extension portion 192. For example, the extension portion 192 may be Figure 8 As shown in the dotted line area, the extension portion 192 is in contact with the cover 180 to support the cover 180 .

[0103] like Figure 8 As shown, two partitions 191 located in the middle are provided with extensions 192, and a partition 191 without the extension 192 is provided between the two partitions 191. If the cover 180 is deformed, the gap between the cover 180 and the partition block 191 may expand to a degree that the target cells can pass through, causing the target cells to enter the liquid outlet groove 140. In this embodiment, the cover 180 is supported by the extension 192 to avoid deformation of the cover 180 and prevent the target cells from flowing out from between the cover 180 and the partition 190, so that the cell screening chip can work normally.

[0104] Along the flow direction of the liquid inlet groove 120, the partitions 191 provided with the extensions 192 and the partitions 191 without the extensions 192 may be arranged alternately. That is, the partitions 191 provided with the extensions 192 and the partitions 191 without the extensions 192 are arranged alternately. The extensions 192 may also be arranged only on one or more partitions 191 at the middle position, and the number and arrangement of the extensions 192 are not limited here.

[0105] Continue to refer to Figures 1 to 3 A liquid outlet groove 140 is provided on both sides of the liquid inlet groove 120. In the two screening arrays 130 located on the opposite sides of the liquid inlet groove 120, the inlet end of the accommodating cavity 131 in one of the screening arrays 130 is staggered with the inlet end of the accommodating cavity 131 in the other screening array.

[0106] Reference Fig. 9 , the accommodating chambers 131 located on both sides of the liquid inlet groove 120 are staggered, so that the liquid inlet groove 120 is staggered and shunted, so that the shunting length of the liquid inlet groove 120 is increased. That is, the liquid inlet groove 120 is shunted throughout, which improves the shunting efficiency. The target cells flow through one accommodating chamber 131 each time, which reduces the probability that the target cells miss the accommodating chamber 131 and improves the capture rate of the target cells. In addition, since the flow of the sample solution in the liquid inlet groove 120 is roughly laminar flow, the flow rate of the sample solution close to the center line of the liquid inlet groove 120 is large, and the flow rate of the sample solution away from the center line of the liquid inlet groove 120 is small, and the accommodating chamber 131 is shunted laterally, providing a lateral flow rate. Since the target cells are affected by the lateral flow rate of one accommodating chamber 131 each time, the target cells flow into the accommodating chamber 131, which improves the capture rate of the target cells.

[0107] Continue to refer to Fig. 9 In each screening unit, the angle between the flow direction of the accommodating cavity 131 and the flow direction of the liquid inlet groove 120 is an acute angle, such as Fig. 9The arrow direction shown is the flow direction of the sample solution in the cell screening chip, where a is an acute angle, for example, the angle of a is 45°. This arrangement allows the sample solution in the inlet groove 120 to flow smoothly into the accommodating cavity 131 in the screening unit, reducing vortices and backflows in the sample solution, thereby reducing the target cells from being impacted and deformed and squeezed out of the screening channel 132 in the screening unit, trapping the target cells in the accommodating cavity 131, and non-target cells with smaller diameters can flow out of the screening channel 132, thereby improving the capture rate of the target cells.

[0108] As the angle a between the flow direction of the accommodating cavity 131 and the flow direction of the inlet groove 120 decreases, the length of the screening unit increases, that is, the flow channel between the inlet groove 120 and the outlet groove 140 for the sample solution to flow through becomes longer. In order to prevent the accumulation of non-target cells with smaller diameters in the screening channel 132 and affect the flow rate of the sample solution, a buffer groove 133 is formed at the outlet end of the screening channel 132, that is, the screening channel 132 is connected to the buffer groove 133.

[0109] Reference Fig.10 and Fig.11 , the buffer tank 133 and the screening channel 132 are projected onto a plane perpendicular to the flow direction of the screening channel 132, and the projection area of ​​the orthographic projection of the buffer tank 133 is larger than the projection area of ​​the orthographic projection of the screening channel 132. With such a configuration, the length of the screening channel 132 can be shortened, the accumulation of non-target cells in the screening channel 132 can be avoided, and the screening channel 132 can be kept unobstructed. In addition, since the sample solution flowing out of the screening channel 132 located upstream passes through the liquid outlet groove 140 and then flows into the screening channel 132 located downstream, it backwashes with the sample solution flowing out of the screening channel 132, resulting in the accumulation of non-target cells in the screening channel 132.

[0110] Continue to refer to Figures 1 to 3 Each screening unit of the screening array 130 may include a accommodating cavity 131 and a screening channel 132. The flow direction of the accommodating cavity 131 coincides with the flow direction of the screening channel 132, which can improve the uniformity of the flow of the sample solution in the screening unit.

[0111] Among the multiple screening units of the screening array 130, at least one screening unit may include a accommodating cavity 131 and at least two screening channels 132. Such a configuration increases the screening path of the sample solution. Compared with the screening unit with only one screening channel 132, the screening unit with multiple screening channels 132 has more diversion paths and increases the diversion flow rate, which can increase the flow rate of the sample solution, shorten the screening time of the sample solution, and improve the screening efficiency.

[0112] In some possible examples, each screening unit includes a containing cavity 131 and two screening channels 132. Fig.12 and Fig.13 The inlet end of each screening channel 132 is connected to the outlet end of the accommodating chamber 131, and the outlet end of each screening channel 132 is connected to the liquid outlet groove 140, and the angle between the flow direction of each screening channel 132 and the flow direction of the liquid outlet groove 140 is an acute angle. Fig.13 As shown by the middle arrow, when the sample solution in the screening channel 132 converges in the liquid outlet groove 140, the sample solution in the liquid outlet groove 140 is prevented from generating a reverse impact on the sample solution flowing out of the screening channel 132 to cause flow obstruction, or even flushing the target cells away from the accommodating chamber 131, thereby improving the capture rate of the accommodating chamber 131 for the target cells.

[0113] In some other possible examples, in each screening array 130, a portion of the screening units include two screening channels 132, and another portion of the screening units include three screening channels 132, further increasing the number of screening channels 132 for diversion, thereby improving the screening efficiency. Fig.14 and Fig.15 Along the extension direction of the liquid inlet groove 120, the screening unit containing three screening channels 132 and the screening unit containing two screening channels 132 are arranged alternately in sequence.

[0114] The widths of the accommodating cavities 131 may be consistent or inconsistent. When the screening array 130 is provided with accommodating cavities 131 of various widths, the accommodating cavities 131 of different widths may capture target cells of different diameters. In a possible example, the width of the accommodating cavities 131 located in the middle of the screening array 130 is greater than the width of the accommodating cavities 131 located at the two ends of the screening array 130, and the widths of the other accommodating cavities 131 may be the same, which is the middle value of the widths of the two accommodating cavities 131.

[0115] In another possible example, the width of the accommodating cavity 131 gradually decreases from the middle to the end of the screening array 130. Fig.16 In the five accommodating cavities 131 shown by the middle dotted lines, the width of the accommodating cavities 131 gradually increases from left to right, i.e., along the flow direction of the inlet groove 120. This arrangement prevents the target cells from accumulating in the accommodating cavities 131 at the ends of the screening array 130, and improves the utilization rate of the accommodating cavities 131 in the middle.

[0116] The liquid inlet groove 120 and the liquid outlet groove 140 may both be serpentine-shaped, and the liquid inlet groove 120 includes at least two linear channels 121 parallel to each other. Fig.17As shown, in the embodiment of the present disclosure, the liquid inlet groove 120 includes eight straight channels 121, and in every two adjacent straight channels 121, the outlet of one straight channel 121 is connected to the inlet of the first curved channel 122, and the outlet of the first curved channel 122 is connected to the inlet of another straight channel 121. With such a configuration, the liquid inlet groove 120 and the liquid outlet groove 140 can be folded to reduce the length of the cell screening chip.

[0117] The first arc channel 122 in the inlet groove 120 may be a semicircular arc, connecting two adjacent straight channels 121 for the sample solution to flow through. The screening array 130 may be partially or completely disposed between the inlet groove 120 and the outlet groove 140 .

[0118] For example, Fig.18 As shown, screening arrays 130 are arranged on both sides of the linear channel 121 of the liquid inlet groove 120, and arc-shaped guide plates 122 are arranged on both sides of the first arc-shaped channel of the liquid inlet groove 120 to guide the sample solution. Fig.19 As shown, the screening array 130 is arranged on both sides of the straight channel 121 and the first curved channel 122 of the liquid inlet groove 120. In this way, when the sample solution flows through the first curved channel 122, it turns and, under the centrifugal effect, the target cells gather on the side of the first curved channel 122 away from the center of the arc, and more target cells are captured on this side. Fig. 20 As shown, screening arrays 130 are arranged on both sides of the first arc-shaped channel 122 of the liquid inlet groove 120, and linear guide plates are arranged on both sides of the linear channel 121 of the liquid inlet groove 120, and the target cells are captured by centrifugal action. Such an arrangement can reduce the processing difficulty of the cell screening chip. Exemplarily, the distance between two adjacent accommodating cavities 131 located on the inner side of the first arc-shaped channel 122 is a first distance, and the distance between two adjacent accommodating cavities 131 located on the outer side of the first arc-shaped channel 122 is a second distance. The first distance is smaller than the second distance, and the inlet ends of the accommodating cavities 131 located on both sides of the first arc-shaped channel 122 are staggered. Each accommodating cavity 131 can be connected to two screening channels 132 to improve the screening efficiency.

[0119] One or more structures mentioned above may be arranged between the liquid inlet groove 120 and the liquid outlet groove 140. For example, screening arrays 130 are arranged on both sides of part of the linear channels 121 of the liquid inlet groove 120, arc-shaped guide plates 122 are arranged on both sides of the first arc-shaped channels connecting the linear channels 121, screening arrays 130 are arranged on both sides of another part of the linear channels 121 of the liquid inlet groove 120, and screening arrays 130 are also arranged on both sides of the first arc-shaped channels connecting this part of the linear channels 121. Alternatively, screening arrays 130 are arranged between the liquid inlet groove 120 and the liquid outlet groove 140, that is, screening arrays 130 are arranged on both sides of all the linear channels 121 and all the first arc-shaped channels 122 of the liquid inlet groove 120.

[0120] In another possible example, the liquid inlet groove 120 and the liquid outlet groove 140 may both be wavy lines. Fig.21 The liquid inlet groove 120 includes at least two second arc-shaped channels 124 connected in sequence, and a screening array 130 is disposed on the side of each second arc-shaped channel 124. In other words, no straight channel is disposed in the liquid inlet groove 120, and the centrifugal effect can be fully utilized, so that the side of each second arc-shaped channel 124 away from the center of each arc can capture the target cells, thereby reducing the accumulation of target cells.

[0121] The wavy line shape can be formed by connecting at least two semicircles in sequence. When the wavy line shape is formed by connecting two semicircles, both the liquid inlet channel 120 and the liquid outlet channel 140 are S-shaped. The wavy line shape can also be a sine curve or a cosine curve.

[0122] In another possible example, the liquid inlet groove 120 and the liquid outlet groove 140 may both be linear. Fig. 22 , the width of the inlet groove 120 gradually increases from the middle of the inlet groove 120 to the end of the inlet groove 120. For example, the width of the inlet groove 120 changes linearly from the end to the middle of the inlet groove 120. In this way, with the plane perpendicular to the center line of the inlet groove 120 as the cross section, the cross-sectional area of ​​the middle of the inlet groove 120 is smaller than the cross-sectional area of ​​the end of the inlet groove 120, the flow rate in the middle of the inlet groove 120 is greater than the flow rate at the end, and the pressure in the middle of the inlet groove 120 is greater than the pressure at the end, so that the target cells in the sample solution enter the screening unit in the middle, and the utilization rate of the screening unit in the middle is improved.

[0123] In another possible example, both the liquid inlet groove 120 and the liquid outlet groove 140 are spiral-shaped. Fig.23, the inlet groove 120 is engaged with the outlet groove 140, and the outlet groove 140 is located on the side of the inlet groove 120 away from the spiral center, that is, the screening array 130 is set on one side of the inlet groove 120. In this way, when the sample solution flows in the spiral inlet groove 120, the target cells flow close to the screening array 130 in the inlet groove 120, and the target cells in the sample solution flow to the screening array 130 under the action of centrifugal force, thereby improving the capture rate of the screening array 130. At the same time, it can also avoid the situation where the target cells are accumulated in some screening units of the screening array 130, while there are no target cells in other screening units, so as to improve the utilization rate of the screening array 130.

[0124] Continue to refer to Figures 1 to 3 The chip body 100 is provided with an injection port 110, which is connected to a liquid inlet groove 120. The sample solution flowing into the liquid inlet groove 120 is a diluted blood sample, that is, the blood sample and the diluent need to be fully mixed first, and then enter the liquid inlet groove 120 through the injection port 110 to separate the target cells.

[0125] The chip body 100 may also be provided with a first injection port 111 and a second injection port 112 at the same time, that is, the chip body 100 adopts a dual injection port. Fig.24 The injection port 110 includes a first injection port 111 and a second injection port 112. The first injection port 110 can be used for blood injection, or other liquid injection such as a fixative, a staining solution, etc., that is, the first injection port 110 is a multi-purpose port. The second injection port 112 is used for diluent injection, that is, the second injection port 110 is a dedicated port.

[0126] The first injection port 111 and the second injection port 112 are respectively connected to the liquid inlet channel 120, and the blood sample entering through the first injection port 111 and the diluent entering through the second injection port 112 flow and mix in the liquid inlet channel 120. The dilution ratio of the blood sample is controlled by controlling the flow rate of the blood sample and the diluent, and the required sample solution is formed in the liquid inlet groove 120. With this arrangement, there is no need to pre-dilute the blood sample, which shortens the detection time of the target cells and improves the detection efficiency.

[0127] When the liquid inlet groove 120 and the liquid outlet groove 140 adopt the above spiral structure, the injection port 110 may also include a first injection port 111 and a second injection port 112. Fig.25, the first injection port 111 and the second injection port 112 are connected to the liquid inlet groove 120 respectively, the first injection port 111 is arranged on the side of the liquid inlet groove 120 close to the screening array 130, and the second injection port 112 is arranged on the side of the liquid inlet groove 120 far away from the screening array 130. With such arrangement, when the blood sample entering from the first injection port 111 and the diluent entering from the second injection port 112 just enter the liquid inlet channel 120, the blood sample and the diluent are not completely mixed. The blood sample is close to the screening array 130, and the diluent is far away from the screening array 130. The blood sample and the diluent interact with each other, and the diluent squeezes the blood sample into the screening array 130, which facilitates the screening array 130 to capture the target cells in the blood sample.

[0128] A liquid inlet connection pipe 160 is also provided between the sample inlet 110 and the liquid inlet groove 120. The liquid inlet connection pipe 160 is used for flow diversion, so that the sample inlet 110 can be connected to multiple liquid inlet grooves 120. On the one hand, the flow rate of the sample solution in the chip body 10 can be increased, thereby shortening the detection time; on the other hand, the screening arrays corresponding to the multiple liquid inlet channels 120 can capture the target cells in parallel, thereby improving the detection efficiency.

[0129] Along the direction from the injection port 110 to the liquid inlet groove 120, the liquid inlet connecting pipe 160 includes at least two levels of liquid inlet diversion pipes arranged in sequence, and each level of liquid inlet diversion pipes includes at least two liquid inlet diversion pipes 161 arranged in parallel. Each liquid inlet diversion pipe 161 located in the upper level is connected with at least two liquid inlet diversion pipes 161 located in the lower level, the first level of liquid inlet diversion pipe close to the injection port 110 is connected with the injection port 110, and the first level of liquid inlet diversion pipe close to the liquid inlet groove 120 is connected with the liquid inlet groove 120 respectively.

[0130] The upper level refers to the level of the adjacent two-level inlet diversion pipeline close to the injection port 110, that is, the level located upstream along the flow direction of the sample solution. The lower level refers to the level of the adjacent two-level inlet diversion pipeline close to the inlet groove 120, that is, the level located downstream along the flow direction of the sample solution.

[0131] In one possible example, refer to Fig.26 The liquid inlet connecting pipe 160 includes two-stage liquid inlet diversion pipes. For the convenience of description, the two-stage liquid inlet diversion pipes are defined as a first-stage liquid inlet diversion pipe 162 and a second-stage liquid inlet diversion pipe 163. Among them, the first-stage liquid inlet diversion pipe 162 is located at Fig.26 The left side is the upper level of the two-level liquid inlet diversion pipeline. The second-level liquid inlet diversion pipeline 163 is located at Fig.26 The right side shown is the next stage in the two-stage liquid inlet diversion pipeline.

[0132] The first-stage liquid inlet diversion pipeline 162 includes two liquid inlet diversion pipelines 161 arranged in parallel, and the inlet ends of the two liquid inlet diversion pipelines 161 are connected and connected to the injection port 110. The outlet ends of the two liquid inlet diversion pipelines 161 are not connected. The second-stage liquid inlet diversion pipeline 163 includes four liquid inlet diversion pipelines 161 arranged in parallel, which can be divided into two groups. One group includes a Fig.26 The inlet ends of the two upper liquid inlet shunt pipes 161 are connected, and the two liquid inlet shunt pipes 161 are connected to the outlet end of one of the two liquid inlet shunt pipes 161 of the first-stage liquid inlet shunt pipe 162. Fig.26 The inlet ends of the two lower inlet diversion pipes 161 are connected, and the two inlet diversion pipes 161 are connected to the outlet end of the other of the two inlet diversion pipes 161 of the first stage inlet diversion pipe 162. The outlet ends of the four inlet diversion pipes 161 of the second stage are connected to one inlet groove 120 respectively.

[0133] Each liquid inlet shunt pipe 161 in the previous stage is connected to two liquid inlet shunt pipes 161 in the next stage, and the cross-sectional area of ​​the liquid inlet shunt pipe 161 in the next stage is half of the cross-sectional area of ​​the liquid inlet shunt pipe 161 in the previous stage, taking the plane perpendicular to the flow direction of the liquid inlet shunt pipe 161 as the cross section. Such an arrangement can improve the uniformity of the flow of the sample solution in the liquid inlet groove 120, so that the pressure in each liquid inlet groove 120 is balanced, and avoid excessive pressure in a certain liquid inlet groove 120, which causes the chip body 100 to be damaged and unable to work normally.

[0134] A liquid outlet connecting pipe 170 is further provided between the liquid outlet groove 140 and the sample outlet 150. The liquid outlet connecting pipe 170 is used for converging the sample solutions of the multiple liquid outlet grooves 140 to the sample outlet 150. Fig.26 , along the direction from the sample outlet 150 to the liquid outlet groove 140, the liquid outlet connecting pipe 170 includes at least two levels of liquid outlet diversion pipes arranged in sequence. Each level of liquid outlet diversion pipe includes at least two liquid outlet diversion pipes 171 arranged in parallel, and each liquid outlet diversion pipe 171 located in the previous level is connected to at least two liquid outlet diversion pipes 171 located in the next level. The first-level liquid outlet diversion pipe close to the sample outlet 150 is connected to the sample outlet 150, and the first-level liquid outlet diversion pipe close to the liquid outlet groove 140 is respectively connected to the liquid outlet groove 140. The specific structure of the liquid outlet connecting pipe 170 can be referred to Fig.26 The structure of the liquid inlet connecting pipe 160 will not be described in detail here.

[0135] The chip body 100 may be provided with only the liquid inlet connecting pipe 160, or only the liquid outlet connecting pipe 170, or both the liquid inlet connecting pipe 160 and the liquid outlet connecting pipe 170, and the number of stages of the liquid inlet connecting pipe 160 may be the same as or different from the number of stages of the liquid outlet connecting pipe 170. When the number of stages of the liquid inlet connecting pipe 160 and the number of stages of the liquid outlet connecting pipe 170 in the chip body 100 are the same, the flow uniformity of the sample solution is better.

[0136] In the cell screening chip provided by the embodiment of the present disclosure, the chip body 100 includes a liquid inlet groove 120 and a liquid outlet groove 140, and a screening array 130 is formed in the partition 190 between the liquid inlet groove 120 and the liquid outlet groove 140. The screening array 130 includes a plurality of screening units, each screening unit includes a accommodating cavity 131 and a screening channel 132, the liquid inlet groove 120, the accommodating cavity 131, the screening channel 132 and the liquid outlet groove 140 are connected in sequence, and the sample solution containing target cells flows from the liquid inlet groove 120 into the accommodating cavity 131 and the screening channel 132, and flows out from the liquid outlet groove 140; the separation of target cells and non-target cells is achieved in the screening unit; at the same time, the first surface of the partition 190 is covered with the second surface of the cover 180, and a gap is formed between the cover 180 and the partition 190. The minimum value of the first distance between the area of ​​the second surface of 180 corresponding to the screening channel 132 and the first surface of the partition 190 is smaller than the radius of the target cell, and the maximum value of the first distance is smaller than the diameter of the target cell, which can prevent the target cell from leaking out from the connection between the gap and the screening channel 132; and the maximum value of the second distance between other areas in the second surface of the cover 180 and the first surface of the partition 190 is smaller than the diameter of the target cell, which can prevent the target cell from leaking out from the gap, thereby ensuring that the cell screening chip can work normally. Compared with the prior art in which the cover is in contact with the partition, in the embodiment of the present disclosure, the sample solution can be diverted between the cover 180 and the partition 190, which increases the flow rate of the sample solution, thereby shortening the separation time of the target cells and improving the separation efficiency of the target cells.

[0137] Embodiment 2

[0138] Reference Fig. 27 The embodiment of the present disclosure provides a cell screening system for separating and identifying target cells. The cell screening system includes the above-mentioned cell screening chip 10, a sampling pump 20 and a waste liquid collection device 70. The sampling port of the cell screening chip 10 is connected to the sampling pump 20, and the sampling port is connected to the waste liquid collection device 70. The sampling pump 20 is used to pump the sample solution into the cell screening chip 10, and the cell screening chip 10 is used to capture the target cells, thereby separating the target cells from the sample solution. The waste liquid collection device 70 is used to collect the waste liquid flowing out of the cell screening chip 10.

[0139] The injection pump 20 includes a sample solution pump 22, and the sample solution pump 22 pumps the sample solution into the cell screening chip 10. In one possible example, the sample solution pump 22 includes a blood sample pump and a diluent pump, and the blood sample and the diluent enter the cell screening chip 10 through a double injection port structure to mix, thereby reducing the time required for detection. In another possible example, the sample solution pump 22 pumps in a diluted blood sample. The diluent may be a phosphate buffer saline (PBS).

[0140] The sample injection pump 20 may also include one or more of a surface treatment liquid pump 21, a buffer liquid pump 23, a fixative liquid pump 24, and a dye liquid pump 25. For example, Fig. 27 As shown in FIG. 1 , the sample injection pump 20 includes a treatment liquid pump 21 , a sample solution pump 22 , a buffer solution pump 23 , a fixative solution pump 24 , and a staining solution pump 25 , which respectively pump different liquids into the cell screening chip 10 .

[0141] When the injection pump 20 includes multiple pumps, a reversing valve 30 is provided between the injection pump 20 and the cell screening chip 10. That is, the output end of the injection pump 20 is connected to one end of the reversing valve 30, and the other end of the reversing valve 30 is connected to the injection port of the cell screening chip 10, so that the liquid in each pump enters the cell screening chip 10 in a certain order through the reversing valve 30.

[0142] The surface treatment liquid can be polyvinyl pyrrolidone (PVP for short), which is used to reduce the flow resistance of the sample solution. The buffer solution can be the same as the diluent, which is also PBS. The fixative solution can be a solution containing 4% paraformaldehyde (PFA for short), which is used to fix the target cells. Specifically, the fixative solution can reduce the elasticity of the cells. The cells after being acted upon by the fixative solution are not easily deformed, and the various structures in the cells are also fixed, thereby achieving the fixation of the cells themselves. The staining solution can be a fluorescent stain, which is used to stain the target cells for easy identification. For example, when the target cells are circulating tumor cells, the fluorescent stain can include a CD with a fluorescent dye. 45 , 4',6-diamidino-2-phenylindole (DAPI) and epithelial cell adhesion molecule (EpCAM) with another fluorescent dye. 45 It is used to mark white blood cells, EpCAM is used to mark circulating tumor cells, and DAPI is used to mark cell nuclei. Target cells can be further identified by staining different cells.

[0143] Continue to refer to Fig. 27 The cell screening system further includes a light source 40, an image acquisition device 50 and a data processing device 60. The light source 40 is used to illuminate the cell screening chip 10 when the image acquisition device 50 is working. The light source 40 can be an LED lamp, or an incandescent lamp or a neon lamp, etc., to provide background light. The light source 40 and the image acquisition device 50 can be located on the same side of the cell screening chip 10, or on both sides of the cell screening chip 10. For example, in this embodiment, the light source 40 is located on the lower side of the cell screening chip 10, and the image acquisition device 50 is located on the upper side of the cell screening chip 10.

[0144] The image acquisition device 50 is signal-connected to the data processing device 60, and is used to acquire images of the cell screening chip 10 and transmit them to the data processing device 60. The image acquisition device can be a charge-coupled device (CCD for short), and the data processing device 60 can be a computer for identifying the number of target cells.

[0145] Continue to refer to Fig. 27 The cell screening system may further include a carrier 80 for placing the cell screening chip 10. The carrier 80 may be a conveyor belt, so that the cell screening chip 10 moves relative to the image acquisition device 50 at a certain speed, thereby ensuring that the image acquisition device 50 can acquire the image of the entire cell screening chip 10.

[0146] In the embodiment of the present disclosure, the cell screening system includes an injection pump 20, a waste liquid collection device 70 and the above-mentioned cell screening chip 10. The injection port of the cell screening chip 10 is connected to the injection pump 20, and the outlet of the cell screening chip 10 is connected to the waste liquid collection device 70. The cell screening system includes the above-mentioned cell screening chip, and thus has the advantages of short separation time and high separation efficiency of target cells. The specific effects are referred to above and will not be repeated here.

[0147] Embodiment 3

[0148] Reference Fig.28 The embodiment of the present disclosure provides a cell screening method, which is applicable to the above-mentioned cell screening system and is used to separate and identify target cells. The cell screening method includes:

[0149] S101 , injecting surface treatment solution and buffer solution into the cell screening chip 10 in sequence for pretreatment.

[0150] The surface treatment liquid is pumped into the cell screening chip 10 through the injection pump 20, and flows out from the waste liquid collection device 70 to perform surface treatment on the functional area of ​​the cell screening chip 10 for capturing target cells. The surface treatment liquid can be PVP. Then, the buffer is pumped into the cell screening chip 10 through the injection pump 20 to rinse the surface treatment liquid, fill the cell screening chip 10, and discharge bubbles.

[0151] S102 , injecting a sample solution containing target cells into the cell screening chip 10 , and the cell screening chip 10 captures the target cells.

[0152] In the embodiments of the present disclosure, the diluted blood sample can be pumped into the cell screening chip 10 by the injection pump 20, that is, the mixed sample solution can be pumped into the cell screening chip 10, or the blood sample and the buffer solution can be separately pumped into the cell screening chip 10 by the injection pump 20 to form a sample solution after mixing.

[0153] The cell screening chip 10 is used to separate target cells from a sample solution, and smaller non-target cells flow out of the cell screening chip 10, while larger target cells are captured and retained by the cell screening chip 10, thereby separating the target cells from the non-target cells. The cell screening chip 10 is the cell screening chip 10 described above, and the separation time of the target cells is short and the separation efficiency is high.

[0154] S103 , sequentially injecting a buffer solution, a fixative solution, a buffer solution, a staining solution, and a buffer solution into the cell screening chip 10 to fix and stain the target cells.

[0155] In the embodiments of the present disclosure, a buffer solution is pumped into the cell screening chip 10 by the injection pump 20 to clean the cell screening chip 10; a fixative is pumped into the cell screening chip 10 by the injection pump 20 to fix the target cells, and the fixative may be PFA; a buffer solution is pumped into the cell screening chip 10 by the injection pump 20 to clean the cell screening chip 10 again to remove the fixative; a staining solution is pumped into the cell screening chip 10 by the injection pump 20 to mark the target cells and further distinguish the cell types in the cell screening chip 10, and the staining solution may be a fluorescent stain.

[0156] For example, the sample solution contains circulating tumor cells, red blood cells, platelets and white blood cells. The diameter of circulating tumor cells is about 10-20 μm, the diameter of red blood cells is about 6-9 μm, the diameter of platelets is about 1-4 μm, and the diameter of white blood cells is about 7-20 μm. When the target cells are circulating tumor cells, the cell screening chip 10 will capture circulating tumor cells and some white blood cells. In order to distinguish the above two types of cells, the two cells can be marked with different fluorescent colors by dyeing liquid to facilitate the identification of circulating tumor cells.

[0157] After staining, the injection pump 20 pumps the buffer solution into the cell screening chip 10 , and the cell screening chip 10 is cleaned again to rinse the staining solution clean, so as to facilitate the subsequent acquisition of the fluorescent image of the cell screening chip 10 .

[0158] S104, the image acquisition device 50 acquires the image of the cell screening chip 10, and transmits the image to the data processing device 60, and the data processing device 60 identifies the target cells.

[0159] In the embodiment of the present disclosure, when the image acquisition device 50 acquires the fluorescent image of the cell screening chip 10, the light source 40 is turned on to provide background light for the cell screening chip 10 and to provide fill light for the image acquisition device 50, so that the image acquisition device 50 can acquire a clearer image. The image acquisition device 50 is connected to the data processing device 60 by signal, and the image is transmitted to the data processing device 60, and the data processing device 60 identifies the number of target cells.

[0160] In the embodiment of the present disclosure, the cell screening chip 10 is injected with a surface treatment solution and a buffer solution for pretreatment in sequence, and then a sample solution is injected, and the target cells in the sample solution are captured by the cell screening chip 10. Since the cell screening method is a method corresponding to the above-mentioned cell screening system, it can shorten the separation time of the target cells and improve the separation efficiency of the target cells, which will not be described in detail here. At the same time, the target cells are fixed with a fixing solution, and the target cells are stained and identified with a staining solution to distinguish them from non-target cells, which is convenient for the identification of the target cells.

[0161] Those skilled in the art should understand that, in the disclosure of the present invention, the orientation or position relationship indicated by the terms "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. are based on the orientation or position relationship shown in the drawings, which are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred system or element must have a specific orientation, be constructed and operated in a specific orientation. Therefore, the above terms should not be understood as limiting the present invention.

[0162] In the description of this specification, the description of reference terms such as "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials, or characteristics described in conjunction with the embodiments or examples are included in at least one embodiment or example of the present disclosure. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0163] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present disclosure, rather than to limit them. Although the present disclosure has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present disclosure.

Claims

1. A cell screening chip, characterized in that: The chip body comprises a liquid inlet groove and a liquid outlet groove, and a partition formed between the liquid inlet groove and the liquid outlet groove, wherein a screening array is formed in the partition, and the screening array comprises a plurality of screening units arranged along the extension direction of the liquid inlet groove; Each of the screening units comprises a receiving chamber and a screening channel, wherein the inlet end of the receiving chamber is communicated with the liquid inlet groove, the outlet end of the receiving chamber is communicated with the inlet end of the screening channel, and the outlet end of the screening channel is communicated with the liquid outlet groove; The width of the accommodating cavity is greater than the diameter of the target cell, and the width of the screening channel is less than the diameter of the target cell; the angle between the flow direction of the accommodating cavity and the flow direction of the liquid inlet groove is an acute angle; along the direction from the middle to the end of the screening array, the width of the accommodating cavity gradually decreases; The second surface of the cover is disposed on the first surface of the partition, and in the second surface of the cover, the minimum value of the first distance between the area corresponding to the screening channel and the first surface of the partition is smaller than the radius of the target cell, and the maximum value of the first distance is smaller than the diameter of the target cell; the maximum value of the second distance between the other areas in the second surface of the cover and the first surface of the partition is smaller than the diameter of the target cell, so that the sample solution can be diverted between the cover and the partition, while preventing the target cell from flowing out from between the cover and the partition; The second surface of the cover is a plane; Along the flow direction of the liquid inlet groove, the end of the liquid inlet groove is closed, a liquid outlet groove is respectively arranged on both sides of the liquid inlet groove, and a screening array is arranged between the liquid inlet groove and each of the liquid outlet grooves; In the two screening arrays, the inlet end of the accommodating cavity in one of the screening arrays is staggered with the inlet end of the accommodating cavity in the other screening array; In each of the plurality of screening units included in the screening array, each of the screening units includes a containing cavity and two screening channels.

2. The cell screening chip according to claim 1, characterized in that: The area of ​​the second surface of the cover corresponding to the screening channel includes a protrusion, the minimum value of the first distance between the protrusion and the first surface of the partition is less than or equal to half the width of the screening channel, the maximum value of the first distance is less than or equal to the width of the screening channel; the maximum value of the second distance is less than or equal to the width of the screening channel.

3. The cell screening chip according to claim 1, characterized in that: The partition part includes a plurality of partition blocks arranged along the flow direction of the liquid inlet groove, a screening unit is formed between adjacent partition blocks, and at least one of the plurality of partition blocks is provided with an extension part, and the extension part is connected to the sealing cover.

4. The cell screening chip according to claim 3, characterized in that: Along the flow guiding direction of the liquid inlet groove, the partition blocks provided with the extension parts and the partition blocks not provided with the extension parts are arranged at intervals.

5. A cell screening system, characterized in that: It comprises an injection pump, a waste liquid collection device and a cell screening chip as described in any one of claims 1 to 4, wherein the injection port of the cell screening chip is connected to the injection pump, and the outlet of the cell screening chip is connected to the waste liquid collection device.

6. A cell screening method, characterized in that: Using the cell screening system according to claim 5, the cell screening method comprises: The cell screening chip is sequentially injected with a surface treatment solution and a buffer solution for pretreatment; injecting a sample solution containing target cells into a cell screening chip, wherein the cell screening chip captures the target cells; injecting a buffer solution, a fixative solution, a buffer solution, a staining solution, and a buffer solution into the cell screening chip in sequence to fix and stain the target cells; The image acquisition device acquires the image of the cell screening chip and transmits the image to the data processing device, and the data processing device identifies the target cells.

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