CN112210474A

CN112210474A - Cell screening chip, cell screening system and method thereof

Info

Publication number: CN112210474A
Application number: CN202011182425.0A
Authority: CN
Inventors: 杨旸; 庄紫云; 康磊
Original assignee: Shanghai Yinghui Medical Instrument Co ltd
Current assignee: Shanghai Yinghui Medical Instrument Co ltd
Priority date: 2020-10-29
Filing date: 2020-10-29
Publication date: 2021-01-12
Also published as: WO2022088965A1

Abstract

The disclosure provides a cell screening chip, a cell screening system and a method thereof, relates to the technical field of biological detection, and is used for solving the technical problem of low capture rate of target cells. The chip body of the cell screening chip is provided with a liquid inlet groove and a liquid outlet groove, the tail end of the liquid inlet groove is closed along the flow guide direction of the liquid inlet groove, and a screening array is formed between the side walls of the liquid inlet groove and the liquid outlet groove which are close to each other; each screening unit of the screening array comprises an accommodating cavity and a screening channel communicated with the accommodating cavity, the accommodating cavity is communicated with the liquid inlet groove, and the screening channel is communicated with the liquid outlet groove; the diameter of the target cell is larger than the width of the screening channel and smaller than the width of the accommodating cavity; the contained angle between the water conservancy diversion direction in holding chamber and the water conservancy diversion direction of feed liquor slot is the acute angle, and the sample solution in the feed liquor slot steadily flows into the holding chamber, reduces the swirl and the backward flow of sample solution, reduces the target cell and receives the impact and the probability that the screening passageway was extruded in deformation, improves the capture rate of target cell.

Description

Cell screening chip, cell screening system and method thereof

Technical Field

The present disclosure relates to the field of biological detection technologies, and in particular, to a cell screening chip, a cell screening system, and a method thereof.

Background

In the field of biological detection, it is often necessary to isolate target cells to facilitate further observation or examination of the target cells, for example, isolation and detection of circulating tumor cells. Circulating Tumor Cells (CTCs) are Tumor cells that are derived from a primary Tumor and enter the blood circulation system, and can develop into metastases through the blood circulation system to propagate to form a secondary Tumor, so that timely separation and detection of the cells are of great significance for monitoring treatment and recurrence of the Tumor.

There are two main methods for screening target cells, one of which is an immunoaffinity capture method, in which the target cells are adsorbed and captured by utilizing the affinity of a specific antigen of the target cells and a specific antibody or aptamer immobilized on a substrate surface or magnetic beads, thereby separating the target cells. Another method is a physical property screening method, which uses the difference of the physical properties of the target cell and other cells to separate the target cell, for example, a microfilter with a channel size smaller than the diameter of the target cell is designed to capture the larger target cell, and the smaller other cells flow out with the buffer solution to separate the target cell.

However, the former method has low capture rate of target cells due to heterogeneity of specific antigens in different types of target cells and even deletion of such specific antigens in some target cells; in the latter method, the target cells are easily broken as the buffer solution flows through the microfilter, resulting in a low capture rate of the target cells.

Disclosure of Invention

In view of the above problems, embodiments of the present disclosure provide a cell screening chip, a cell screening system, and a method thereof for improving a capture rate of target cells.

In order to achieve the above object, the embodiments of the present disclosure provide the following technical solutions:

in a first aspect, an embodiment of the present disclosure provides a cell screening chip, including a chip body, where the chip body is provided with a liquid inlet groove and a liquid outlet groove, a terminal of the liquid inlet groove is closed along a flow guide direction of the liquid inlet groove, and a screening array is formed between side walls of the liquid inlet groove and the liquid outlet groove that are close to each other; the screening array comprises a plurality of screening units, each screening unit comprises an accommodating cavity and a screening channel, the inlet end of the accommodating cavity is communicated with the liquid inlet groove, the outlet end of the accommodating cavity 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 larger than the diameter of the target cell, and the width of the screening channel is smaller than the diameter of the target cell; in each screening unit, an included angle between the flow guide direction of the accommodating cavity and the flow guide direction of the liquid inlet groove is an acute angle.

The cell screening chip provided by the embodiment of the disclosure has the following advantages:

in the cell screening chip provided by the embodiment of the disclosure, the chip body is provided with the liquid inlet groove and the liquid outlet groove, and the tail end of the liquid inlet groove is sealed along the flow guide direction of the liquid inlet groove. A screening array is arranged between the side walls of the liquid inlet groove and the liquid outlet groove, which are close to each other, namely the screening array is positioned on the side surface of the liquid inlet groove. The screening array comprises a plurality of screening units, each screening unit comprises a containing cavity and a screening channel, the liquid inlet grooves, the containing cavities, the screening channels and the liquid outlet grooves are communicated in sequence, and sample solution containing target cells flows into the containing cavities and the screening channels in sequence through the liquid inlet grooves and flows out through the liquid outlet grooves. Because the width of the accommodating cavity is larger than the diameter of the target cell, the width of the screening channel is smaller than the diameter of the target cell, the target cell cannot be intercepted in the accommodating cavity through the screening channel, and the non-target cell with the diameter smaller than the width of the screening channel flows out of the screening channel, so that the separation of the target cell and the non-target cell is realized. Simultaneously, because in every screening unit, the contained angle between the water conservancy diversion direction of holding chamber and the water conservancy diversion direction of feed liquor slot is the acute angle for in the sample solution in the feed liquor slot steadily flows into the holding chamber, reduce swirl and backward flow in the sample solution, thereby reduce target cell and receive the impact and deformation extrusion screening passageway, improved target cell's entrapment rate, and then improved target cell's capture rate.

In a second aspect, an embodiment of the present disclosure further provides a cell screening system, which includes a sample pump, a waste liquid collecting device, and the cell screening chip as described above, wherein a sample inlet of the cell screening chip is connected to the sample pump, and a sample outlet of the cell screening chip is connected to the waste liquid collecting device.

The cell screening system provided by the embodiments of the present disclosure includes the cell screening chip, and thus has the advantage of high capture rate of target cells, and specific effects are referred to above and are not described herein again.

In a third aspect, an embodiment of the present disclosure further provides a cell screening method, which uses the cell screening system, and the cell screening method includes:

sequentially injecting surface treatment liquid and buffer liquid into the cell screening chip for pretreatment; injecting a sample solution containing target cells into a cell screening chip, which captures the target cells; sequentially injecting a buffer solution, a fixing solution, a buffer solution, a staining solution and a buffer solution into the cell screening chip, wherein the buffer solution is used for cleaning the cell screening chip, the fixing solution is used for typing the target cells in the cell screening chip, and the staining solution is used for staining and marking the target cells; the image acquisition device acquires an image of the cell screening chip and transmits the image to the data processing device, and the data processing device identifies the target cell.

The cell screening method provided by the embodiment of the disclosure has the following advantages:

in the embodiment of the present disclosure, the cell screening chip is pretreated and then injected into the sample solution, and the cell screening chip captures the target cells in the sample solution. Meanwhile, the target cells are shaped by using the fixing solution, and are subjected to staining identification by using the staining solution so as to be distinguished from non-target cells, so that the target cells are conveniently identified.

Drawings

In order to more clearly illustrate the embodiments of the present disclosure or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present disclosure, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a schematic diagram of a cell screening chip according to an embodiment of the present disclosure;

fig. 2 is a schematic structural diagram of a chip body in an embodiment of the disclosure;

FIG. 3 is an enlarged view of a portion of FIG. 2 at A;

FIG. 4 is a schematic distribution diagram of receiving cavities of different widths in an embodiment of the present disclosure;

fig. 5 is a schematic flow diagram of a sample solution in a chip body in an embodiment of the disclosure;

FIG. 6 is a schematic diagram of another structure of a cell screening chip according to an embodiment of the present disclosure;

FIG. 7 is an enlarged view of a portion of FIG. 6 at B;

fig. 8 is a schematic view of a first structure of a liquid inlet groove in an embodiment of the disclosure;

FIG. 9 is a first enlarged partial view taken at C in FIG. 8;

FIG. 10 is a second enlarged partial view taken at C in FIG. 8;

FIG. 11 is a third enlarged partial view at C of FIG. 8;

fig. 12 is a schematic view of a second structure of a liquid inlet groove in an embodiment of the disclosure;

fig. 13 is a schematic view of a third structure of a liquid inlet groove in an embodiment of the present disclosure;

fig. 14 is a partial enlarged view at D in fig. 13;

fig. 15 is a schematic view of a fourth structure of a liquid inlet groove in an embodiment of the present disclosure;

FIG. 16 is a schematic diagram of a first structure of a dual injection port in an embodiment of the disclosure;

FIG. 17 is a second schematic diagram of a dual inlet port in an embodiment of the disclosure;

FIG. 18 is a partial view of a second configuration of dual injection ports in an embodiment of the present disclosure;

FIG. 19 is a schematic diagram of an inlet connection pipe in an embodiment of the disclosure;

FIG. 20 is another schematic diagram of an inlet connection pipe in an embodiment of the disclosure;

FIG. 21 is a schematic diagram of a cell screening system in an embodiment of the present disclosure;

fig. 22 is a flow chart of a cell screening method in an embodiment of the present disclosure.

Description of reference numerals:

10. a cell screening chip; 100. A chip body;

110. a sample inlet; 111. A first sample inlet;

112. a second sample inlet; 120. A liquid inlet groove;

121. a linear channel; 122. A first arcuate channel;

123. an arc-shaped guide plate; 124. A second arcuate channel;

130. screening the array; 131. An accommodating cavity;

132. screening a channel; 133. A buffer tank;

140. a liquid outlet groove; 150. A sample outlet;

160. the liquid inlet is connected with a pipeline; 161. A liquid inlet shunting pipeline;

162. a first stage inlet liquid diversion pipeline; 163. A second stage inlet liquid diversion pipeline;

170. a liquid outlet connecting pipeline; 171. A liquid outlet diversion pipeline;

180. sealing the cover; 20. A sample injection pump;

21. a surface treatment liquid pump; 22. A sample solution pump;

23. a buffer pump; 24. Fixing a liquid pump;

25. a dyeing liquid pump; 30. A diverter valve;

40. a light source; 50. An image acquisition device;

60. a data processing device; 70. A waste liquid collection device;

80. a stage; l1, width of the receiving cavity;

l2, width of screening channel; a. And (4) an included angle.

Detailed Description

In the related art, the cell screening chip includes a chip body and a sealing cover, the chip body and the sealing cover are sealed, and a cavity for screening target cells is formed between the chip body and the sealing cover.

In order to solve the technical problem that the capture rate of target cells is low, the cell screening chip in the embodiment of the disclosure comprises a chip body, wherein the chip body is provided with a liquid inlet groove and a liquid outlet groove arranged on the side surface of the liquid inlet groove, and the tail end of the liquid inlet groove is sealed along the flow guide direction of the liquid inlet groove. Be provided with a plurality of screening units between feed liquor slot and the play liquid slot, every screening unit includes holding chamber and screening passageway, the feed liquor slot, the holding chamber, the screening passageway communicates with play liquid slot in proper order, the width in holding chamber is greater than the diameter of target cell, the width of screening passageway is less than the diameter of target cell, and the water conservancy diversion direction in holding chamber and the feed liquor slot in the contained angle between the flow direction of sample solution be the acute angle, make the sample solution in the feed liquor slot steadily get into in the holding chamber, reduce the swirl and the backward flow of sample solution, thereby it extrudes the screening passageway to reduce the target cell and receive the impact, improve the capture rate of target cell.

In order to make the above objects, features and advantages of the embodiments of the present disclosure more comprehensible, embodiments of the present disclosure are described in detail below with reference to the accompanying drawings. It is to be understood that the described embodiments are merely a subset of the disclosed embodiments and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments disclosed herein without making any creative effort, shall fall within the protection scope of the present disclosure.

Example one

In order to separate target cells in a sample solution for subsequent observation or inspection, embodiments of the present disclosure provide a cell screening chip, which may be made of a transparent material and used for subsequent identification of the target cells. Referring to fig. 1, the cell screening chip in the embodiment of the present disclosure includes a chip body 100 and a cap 180, wherein the cap 180 is capped on the chip body for sealing the chip body 100 and preventing the sample solution from overflowing the cell screening chip.

The chip body 100 is provided with a sample inlet 110, a liquid inlet channel 120, a screening array 130 (shown by dotted lines in fig. 2), a liquid outlet channel 140 and a sample outlet 150. Referring to fig. 2, one end of the liquid inlet groove 120 is communicated with the sample inlet 110, and the liquid inlet groove 120 and the liquid outlet groove 140 extend in the same direction. For example, as shown in fig. 2, the liquid inlet groove 120 and the liquid outlet groove 140 may be horizontally disposed.

The end of the liquid inlet groove 120 is closed along the flow guiding direction of the liquid inlet groove 120. For example, the left end of the liquid inlet channel 120 shown in fig. 2 is communicated with the sample inlet 110, and the right end is closed. The screening array 130 is disposed between the side walls of the liquid inlet groove 120 and the liquid outlet groove 140 close to each other, and is communicated with the screening array 130.

In the embodiments of the present disclosure and in the following embodiments, the extending direction of the liquid inlet groove 120 is also the flow guiding direction of the liquid inlet groove 120, and the extending direction of the liquid outlet groove 140 is also the flow guiding direction of the liquid outlet groove 140.

In the embodiment of the present disclosure, 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 is a surface facing the opening of the liquid inlet groove 120, the side wall of the liquid inlet groove 120 is a surface parallel to the extending direction of the liquid inlet groove 120, and the end of the liquid inlet groove 120 is a surface starting or ending in the extending direction of the liquid inlet groove 120. It is understood that the side wall of the liquid inlet groove 120 connects both end portions of the liquid inlet groove 120.

Taking the structure shown in fig. 1 as an example, the left and right surfaces of the liquid inlet groove 120 are the end portions of the liquid inlet groove 120, the front and rear vertical surfaces 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 is not described in detail herein.

The side wall of the liquid inlet groove 120 and the liquid outlet groove 140 close to each other means 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.

The liquid inlet groove 120 and the liquid outlet groove 140 may be first grooves formed in the chip body 100. One end of the liquid outlet groove 140 communicates with the sample outlet 150. The sample solution flows in from the sample inlet 110, and flows to the sample outlet 150 through the liquid inlet channel 120, the screening array 130 and the liquid outlet channel 140 in sequence.

The screening array 130 is used for trapping and capturing target cells and may include a plurality of screening units. Referring to fig. 2 and 3, in the present embodiment, the screening array 130 includes seven screening units. The screening units in the screening array 130 are spaced apart from each other along the extending direction of the liquid inlet channel 120, i.e. the screening units are not communicated with each other. In one possible example, the screening unit is a second groove formed on the chip body 100, and every two adjacent screening units are separated by the chip body 100.

Each screening unit comprises an accommodating cavity 131 and a screening channel 132 communicated with the accommodating cavity 131, and the liquid inlet groove 120, the accommodating cavity 131, the screening channel 132 and the liquid outlet groove 140 are communicated in sequence. That is, the liquid inlet channel 120 is communicated with the inlet end of the accommodating chamber 131, the outlet end of the accommodating chamber 131 is communicated with the inlet end of the screening channel 132, the outlet end of the screening channel 132 is communicated with the liquid outlet channel 140, and the liquid inlet channel 120, the accommodating chamber 131, the screening channel 132 and the liquid outlet channel 140 form a channel for the sample solution to flow through, so that the sample solution flows through the cell screening chip.

In each screening unit, an included angle between the flow guiding direction of the accommodating cavity 131 and the flow guiding direction of the liquid inlet groove 120 is an acute angle, and the flow guiding direction of the accommodating cavity 131 is related to the shape of the accommodating cavity 131. The direction of the arrow shown in FIG. 5 is the direction of flow in the cell screening chip, and a is an acute angle, for example, an angle of 45 degrees. So set up, can make the sample solution in the feed liquor slot 120 smoothly flow into the holding chamber 131 in the screening unit, reduce swirl and backward flow in the sample solution to it extrudes the screening passageway 132 in the screening unit to reduce target cell and receive the impact and deformation. The target cells are trapped in the accommodating cavity 131, and the non-target cells with smaller diameters can flow out from the screening channel 132, so that the capture rate of the target cells is improved.

In the above embodiments, the cover 180 may be located above the chip body 100 or below the chip body 100. Specifically, the cover 180 covers the chip body 100, and a bottom surface of the cover 180 is attached to an upper surface of the chip body 100. Target cells are prevented from overflowing from the gap between the two surfaces to the region for screening the target cells in the chip body 100, the retention rate of the target cells is improved, and the detection accuracy of the cell screening chip is further improved.

The cell screening chip may be a microfluidic chip, and the material of the cell screening chip may be polymethyl methacrylate (PMMA), Polycarbonate (PC), Polystyrene (PS), glass, or the like. That is, the material of the chip body 100 may be PMMA, PC, PS, or glass.

The cover 180 may be of a flat plate construction to facilitate the machining and forming of the cover 180. The bottom surface of the cap 180 may have a size corresponding to that of the upper surface of the chip body 100. As shown in fig. 1, the cap 180 covers the entire upper surface of the chip body 100. The size of the bottom surface of the cap 180 may be smaller than the size of the upper surface of the chip body 100, and the cap 180 covers a portion of the upper surface of the chip body 100. I.e., the functional region of the chip body 100 is covered to reduce the volume of the cover 180. Of course, the cover 180 may have other structures.

The cap 180 may be formed by injection molding, and the cap 180 and the chip body 100 are bonded, for example, by thermal bonding, adhesive bonding, or ultrasonic bonding. The cap 180 may also be a film layer formed on the chip body 100 by a film-attaching process. The cap 180 may also be integrally formed with the chip body 100, for example, the cap 180 is integrally formed with the chip body 100 by 3D printing. The connection between the cap 180 and the chip body 100 may be performed by other methods, which are not described herein.

The chip body 100 may be injection molded. When the chip body 100 is formed by injection molding, a mold for forming the chip body 100 is required, and the mold may be formed by electroforming, machining, or etching. The chip body 100 may be manufactured by other micro-fabrication techniques such as laser etching and photolithography.

In each screening unit, the flow direction of the receiving chamber 131 and the flow direction of the screening channel 132 may be parallel or coincide. So set up, avoid sample solution to take place the turn in the screening unit, reduce sample solution's flow loss. In the present embodiment, the flow direction of the accommodating cavity 131 coincides with the flow direction of the screening channel 132. So set up, can further improve the homogeneity that the sample solution flows in the screening unit. Optionally, the flow guiding directions in the respective screening units are parallel to each other. With this arrangement, the flow direction of each screening unit in the screening array 130 is the same, so as to improve the uniformity of the flow of the sample solution in the screening array 130.

Referring to fig. 3, in each of the sieving units, the width L1 of the receiving chamber 131 is greater than the diameter of the target cell, and the width L2 of the sieving channel 132 is smaller than the diameter of the target cell. Since a plurality of cells in the target cell have different sizes, the diameter of the target cell is generally a range of values. In this and the following examples, the diameter of the target cell means the minimum value of the range of diameters of the target cell, that is, the diameter value of the smallest cell among the target cells. For example, when the target cells are circulating tumor cells, the diameter of the target cells is about 10-20 μm, and the width of the screening channel 132 is less than 10 μm, and may be 8 μm. The width of the accommodating cavity 131 is greater than 10 μm, and may be 20 μm. So configured, the target cells cannot pass through the screening channel 132, so that the target cells are trapped in the accommodating chamber 131. Non-target cells having a diameter smaller than the screening channel 132 can flow out of the screening channel 132, so that the target cells are separated from the sample solution and trapped in the holding chamber 131. Preferably, each receiving chamber 131 captures and retains a single target cell therein.

The receiving cavity 131 may be a receiving groove formed in the chip body 100, and an opening of the receiving groove faces the cover 180. The width of the accommodating cavity 131 refers to the distance between two opposite side walls of the accommodating groove, such as the length of L1 shown in fig. 3. The screening channel 132 may be a flow guide groove or a flow guide hole formed in the chip body 100. When the screening channel 132 is a channel, the opening of the channel is facing the cover 180, and the width of the screening channel 132 refers to the distance between two opposite side walls of the channel, as shown in fig. 3 by the length L2.

Since the width of the receiving cavity 131 is larger than the width of the screening channel 132, the receiving cavity 131 and the screening channel 132 form a step surface facing the liquid inlet groove 120. The step surface and the two side walls of the accommodating cavity 131 and the two side walls of the screening channel 132 may be in arc transition to reduce the flow resistance of the sample solution.

The widths of the plurality of receiving cavities 131 in the screening array 130 may or may not be uniform. When the receiving cavities 131 with various widths are arranged in the screening array 130, the receiving cavities 131 with different widths can capture target cells with different diameters.

To avoid accumulation of multiple target cells in one receiving chamber 131, such as receiving chambers 131 near the ends of the screening array 130, in one possible example, the width of the receiving chamber 131 in the middle of the screening array 130 is greater than the width of the receiving chambers 131 at both ends of the screening array 130. The widths of the other receiving cavities 131 may be the same, which is the middle of the widths of the two receiving cavities 131. So set up, the width that is located the holding chamber 131 in screening array 130 middle part is great, and the target cell of being convenient for gets into to avoid the target cell to pile up in the holding chamber 131 of the tip of screening array 130, improve the utilization ratio that is located the holding chamber 131 in middle part.

In another possible example, the width of the receiving cavity 131 gradually decreases in a direction from the middle to the end of the screening array 130. In the five accommodating cavities 131 from the left end to the middle as shown by the dotted line in fig. 4, the width of the accommodating cavity 131 gradually increases along the flow guiding direction of the liquid inlet groove 120. So set up, be close to screening array 130 middle part more, the width of holding chamber 131 is big more, the more target cell transition of being convenient for to avoid target cell to pile up in the holding chamber 131 of the tip of screening array 130, improve the utilization ratio of the holding chamber 131 at middle part.

The cross-sectional shape of the accommodating cavity 131 may be trapezoidal, quadrangular, U-shaped, parabolic, or semicircular, taking a plane parallel to the upper surface of the chip body 100 as a cross section, and the specific shape of the accommodating cavity 131 is not limited herein. The shapes of the receiving cavities 131 may be the same or different. The width of the outlet end of the accommodating chamber 131 is less than or equal to the width of the inlet end of the accommodating chamber 131 and greater than the diameter of the target cells for trapping and trapping the target cells.

The length of the screening unit increases as the degree of the included angle a between the flow guiding direction of the accommodating cavity 131 and the flow guiding direction of the liquid inlet groove 120 decreases. That is, the flow path between the inlet channel 120 and the outlet channel 140 through which the sample solution flows becomes long. To avoid the accumulation of non-target cells with smaller diameters in the screening channel 132 to affect the flow rate of the sample solution, in one possible example, the length of the receiving chamber 131 may be increased, thereby reducing the length of the screening channel 132.

In another possible example, the outlet end of the screening channel 132 is formed with a buffer groove 133, i.e. the screening channel 132 communicates with the buffer groove 133. The buffer groove 133 and the screening channel 132 are projected to a plane perpendicular to the flow guiding direction of the screening channel 132, and the projection area of the orthographic projection of the buffer groove 133 is larger than that of the orthographic projection of the screening channel 132. With such an arrangement, the length of the screening channel 132 can be shortened, accumulation of non-target cells in the screening channel 132 can be avoided, and the smoothness of the screening channel 132 can be maintained. In addition, since the sample solution flowing out of the upstream screening channel 132 may flow into the downstream screening channel 132 and form a back-wash phenomenon with the sample solution flowing out of the screening channel 132, the buffer tank 133 can reduce or prevent the back-wash phenomenon, thereby preventing the non-target cells from accumulating in the screening channel 132.

In one possible example, the center line of the buffer tank 133 coincides with the center line of the screening channel 132 to improve the uniformity of the flow of the sample solution in the screening unit. Referring to fig. 6 and 7, the outlet end of each screening channel 132 is formed with a buffer groove 133, the opening of the buffer groove 133 faces the liquid outlet groove 140, the buffer groove 133 is roughly shaped like a triangular prism, and as shown by the dotted line in fig. 7, the plane formed by removing the sharp corner at the end part of the left side wall of the screening channel 132 is a part of the groove bottom of the buffer groove 133.

The buffer groove 133 may also have a quadrangular prism shape or a semi-cylindrical shape, and the specific shape of the buffer groove 133 is not limited herein. Wherein, one side of the quadrangular buffer groove 133 is communicated with the outlet end of the screening channel 132, and the arc surface of the semicylindrical buffer groove 133 is communicated with the outlet end of the screening channel 132.

At least one of the two sides of the liquid inlet channel 120 is provided with the liquid outlet channel 140, so that a longer screening array 130 can be arranged between the liquid inlet channel 120 and the liquid outlet channel 140, and the capture rate of the target cells is improved. For example, two liquid outlet grooves 140 are disposed on two sides of the liquid inlet groove 120, and the screening arrays 130 are disposed between two sidewalls of the liquid inlet groove 120 and a sidewall of the liquid outlet groove 140 close to the liquid inlet groove 120. With such an arrangement, the length of the screening array 130 can be further increased, and the screening efficiency of the target cells can be improved.

As shown in fig. 2, the liquid outlet grooves 140 are disposed on both upper and lower sides of the liquid inlet groove 120, a screening array 130 is disposed between the upper sidewall of the liquid inlet groove 120 and the lower sidewall of the liquid outlet groove 140 located on the upper side of the liquid inlet groove 120, and a screening array 130 is disposed between the lower sidewall of the liquid inlet groove 120 and the upper sidewall of the liquid outlet groove 140 located on the lower side of the liquid inlet groove 120.

With continued reference to fig. 2, one end of the liquid outlet groove 140 is communicated with the sample outlet 150, the other end is closed, and both liquid outlet grooves 140 are communicated with the sample outlet 150. That is, the two outlet grooves 140 converge at the outlet 150, and the sample solution after separating the target cells flows out of the cell screening chip through the outlet 150.

In a possible example, the inlet channel 120 and the outlet channel 140 may both have a serpentine shape, the inlet channel 120 comprising at least two rectilinear channels 121 parallel to each other. In the embodiment of the present disclosure, as shown in fig. 8, the liquid inlet groove 120 includes eight straight passages 121. In every two adjacent linear channels 121, the outlet of one of the linear channels 121 is communicated with the inlet of the first arc-shaped channel 122, and the outlet of the first arc-shaped channel 122 is communicated with the inlet of the other linear channel 121. That is, the straight passage 121 communicates with the first arc-shaped passage 122 in sequence. Thus, the inlet channel 120 and the outlet channel 140 can be folded to reduce the length of the cell screening chip.

The flow direction of the linear channel 121 in the liquid inlet groove 120 may be the same as the length direction of the cell screening chip. As shown in FIG. 8, the flow direction of the linear channel 12 is parallel to the longitudinal direction of the cell screening chip. The flow direction of the linear channel 121 in the liquid inlet groove 120 may also be the same as the width direction of the cell screening chip. The number and the flow direction of the linear channels 121 in the liquid inlet groove 120 are arranged according to the use requirement of the cell screening chip.

The first arc-shaped channel 122 in the liquid inlet groove 120 may be a semi-arc, and connects two adjacent straight channels 121 for the sample solution to flow through. The screening array 130 may be partially disposed between the liquid inlet channel 120 and the liquid outlet channel 140, or the screening array 130 may be disposed entirely.

In one possible example, as shown in fig. 9, screening arrays 130 are disposed on both sides of the linear channel 121 of the liquid inlet groove 120, and arc-shaped guide plates 123 are disposed on both sides of the first arc-shaped channel of the liquid inlet groove 120 to guide the sample solution.

In another possible example, as shown in fig. 10, the screening arrays 130 are disposed on both sides of the straight passage 121 and the first arc-shaped passage 122 of the liquid inlet groove 120. So configured, when the sample solution turns when flowing through the first arc-shaped channel 122, the target cells are gathered toward the side of the first arc-shaped channel 122 away from the center of the arc under the centrifugal action, and the side captures more target cells.

In yet another possible example, as shown in fig. 11, the screening arrays 130 are disposed on both sides of the first arc-shaped channel 122 of the liquid inlet groove 120, and the linear baffles are disposed on both sides of the linear channel 121 of the liquid inlet groove 120 to capture the target cells by centrifugation. So set up, can reduce the processing degree of difficulty of cell screening chip. Illustratively, the distance between two adjacent receiving cavities 131 located inside the first arc-shaped channel 122 is a first distance, and the distance between two adjacent receiving cavities 131 located outside 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 at the two sides of the first arc-shaped channel 122 are staggered. Each receiving chamber 131 may communicate with two screening channels 132 to improve screening efficiency.

In another possible example, the liquid inlet channel 120 and the liquid outlet channel 140 may be both wavy lines. Referring to fig. 12, the liquid inlet groove 120 includes at least two second arc-shaped channels 124 communicated with each other in sequence, and a screening array 130 is disposed on a side of each second arc-shaped channel 124. That is, the straight passage is not provided in the liquid inlet groove 120, and the centrifugal effect can be fully utilized, so that the side of each second arc-shaped passage 124 far from the center of each arc-shaped passage catches the target cells, thereby reducing the accumulation of the target cells.

The wavy line shape may be formed by connecting at least two semicircles in sequence, and when the wavy line shape is formed by connecting two semicircles, the liquid inlet groove 120 and the liquid outlet groove 140 are both S-shaped. The wavy line shape may be a sine curve or a cosine curve.

In another possible example, the liquid inlet groove 120 and the liquid outlet groove 140 may be both linear. Referring to fig. 13 and 14, the width of the liquid inlet groove 120 gradually increases toward the end of the liquid inlet groove 120 along the middle of the liquid inlet groove 120. The left end of the first section of the liquid inlet groove 120 shown in fig. 14 is close to the middle of the whole liquid inlet groove 120, the right end of the first section of the liquid inlet groove 120 shown in fig. 14 is close to the end of the whole liquid inlet groove 120, and the width of the first section of the liquid inlet groove 120 shown in fig. 14 gradually increases from left to right.

It can be understood that, along the flow guiding direction of the liquid inlet groove 120, the width of the whole liquid inlet groove 120 is firstly reduced, then increased and then reduced. For example, the width of the liquid inlet groove 120 varies linearly from the end to the middle of the liquid inlet groove 120. In this way, the plane perpendicular to the flow guiding direction of the liquid inlet groove 120 is taken as the cross section, and the cross section area of the middle part of the liquid inlet groove 120 is smaller than that of the end part of the liquid inlet groove 120. The flow velocity in the middle of the liquid inlet channel 120 is greater than that in the end, so that the pressure of the side wall in the middle of the liquid inlet channel 120 is greater than that in the end, thereby promoting the target cells in the sample solution to enter the screening unit in the middle, and improving the capture rate of the screening unit in the middle. So set up, reduce the piling up of target cell in the screening unit of tip, improve the utilization ratio of the screening unit at middle part, target cell can be held back in a plurality of screening units in the screening array by the dispersion, and then makes most screening units all can catch target cell to follow-up discernment target cell.

In another possible example, the liquid inlet groove 120 and the liquid outlet groove 140 are both spiral, and the liquid inlet groove 120 is embedded in the liquid outlet groove 140. Wherein, two sides of the liquid inlet groove 120 can be respectively provided with a liquid outlet groove 140, and the liquid inlet groove 120 can also be provided with a liquid outlet groove 140 on one side.

Illustratively, referring to fig. 15, the liquid outlet groove 140 is located on one side of the liquid inlet groove 120 away from the center of the spiral, that is, the liquid outlet groove 140 is disposed on the outer side of the liquid inlet groove 120, and the screening array 130 is disposed on one side of the liquid inlet groove 120. So configured, when the sample solution flows in the spiral inlet channel 120, the target cells flow close to the screening array 130 in the inlet channel 120, and the target cells in the sample solution flow toward the screening array 130 under the centrifugal force, thereby increasing the capture rate of the screening array 130. Meanwhile, the situation that target cells are accumulated in part of the screening units of the screening array 130 and no target cells exist in other parts of the screening units can be avoided, and the utilization rate of the screening array 130 can be improved due to the arrangement.

With reference to fig. 2, the chip body 100 shown in fig. 2 is provided with a sample inlet 110, the sample inlet 110 is communicated with the liquid inlet channel 120, and the sample solution flowing into the liquid inlet channel 120 is a diluted blood sample. That is, the blood sample and the diluent are mixed sufficiently and then enter the liquid inlet channel 120 through the sample inlet 110 to separate the target cells.

The chip body 100 may also be provided with a first sample inlet 111 and a second sample inlet 112 at the same time, i.e. the chip body 100 adopts dual sample inlets. Referring to fig. 16, the sample inlet 110 may include a first sample inlet 111 and a second sample inlet 112. The first sample inlet 110 can be used for blood sample injection, or other liquid sample injections such as stationary liquid and staining solution, that is, the first sample inlet 110 is a multi-purpose port. The second sample inlet 112 is used for injecting a diluent, i.e., the second sample inlet 110 is a dedicated port.

The first sample inlet 111 and the second sample inlet 112 are respectively communicated with the liquid inlet groove 120, and a blood sample entering from the first sample inlet 111 and a diluent entering from the second sample inlet 112 flow and are mixed in the liquid inlet groove 120. The dilution ratio of the blood sample is controlled by controlling the flow rates of the blood sample and the diluent, and a desired sample solution is formed in the inlet channel 120. According to the arrangement, a blood sample does not need to be diluted in advance, the detection time of the target cells is shortened, and the detection efficiency is improved.

When the liquid inlet groove 120 and the liquid outlet groove 140 take the spiral structure, and the liquid outlet groove 140 is disposed outside the liquid inlet groove 120, the sample inlet 110 may also include the first sample inlet 111 and the second sample inlet 112.

Referring to fig. 17 and 18, the first liquid inlet 111 and the second liquid inlet 112 are respectively communicated with the liquid inlet groove 120. The first sample inlet 111 is disposed on one side of the liquid inlet groove 120 close to the screening array 130, and the second sample inlet 112 is disposed on one side of the liquid inlet groove 120 far from the screening array 130. With this arrangement, when the blood sample entering from the first sample inlet 111 and the diluent entering from the second sample inlet 112 enter the liquid inlet groove 120, the blood sample and the diluent are not completely mixed. The blood sample is close to the screening array 130, the diluent is far away from the screening array 130, the blood sample interacts with the diluent, and the diluent extrudes the blood sample into the screening array 130, so that the screening array 130 can capture target cells in the blood sample conveniently.

An inlet connection pipe 160 may be further disposed between the inlet 110 and the inlet grooves 120, and the inlet connection pipe 160 is used for splitting, so that the inlet 110 may be connected to a plurality of inlet grooves 120. On the one hand, the flow rate of the sample solution in the chip body 100 can be improved, so that the detection time is shortened, on the other hand, the parallel capture of the target cells by the screening arrays corresponding to the plurality of liquid inlet grooves 120 is realized, and the detection efficiency is improved.

Along the direction from the sample inlet 110 to the liquid inlet groove 120, the liquid inlet connecting pipeline 160 includes at least two stages of liquid inlet shunting pipelines arranged in sequence, and each stage of liquid inlet shunting pipeline includes at least two liquid inlet shunting pipelines 161 arranged in parallel. Each liquid inlet shunting pipeline 161 positioned in the upper stage is communicated with at least two liquid inlet shunting pipelines 161 positioned in the lower stage, the first-stage liquid inlet shunting pipeline close to the sample inlet 110 is communicated with the sample inlet 110, and the first-stage liquid inlet shunting pipelines close to the liquid inlet grooves 120 are respectively communicated with the liquid inlet grooves 120.

The upper stage refers to a stage close to the sample inlet 110 in the two adjacent stages of liquid inlet diversion pipelines, that is, a stage located at the upstream along the diversion direction. The next stage refers to a stage close to the liquid inlet groove 120 in the two adjacent stages of liquid inlet branch pipes, that is, a stage located downstream in the flow direction of the sample solution.

In one possible example, referring to fig. 19, the inlet connection conduit 160 comprises a two-stage inlet split conduit. For convenience of description, the two-stage inlet flow-splitting ducts are defined as the first stage inlet flow-splitting duct 162 and the second stage inlet flow-splitting duct 163, respectively. The first-stage liquid inlet diversion pipeline 162 is located on the left side of fig. 19, and is the upper stage of the two-stage liquid inlet diversion pipeline. The second stage feed split pipe 163 is located on the right side as viewed in fig. 19, being the next stage in the two stage feed split pipe.

The first-stage liquid inlet diversion pipeline 162 comprises two liquid inlet diversion pipelines 161 arranged in parallel, and the inlet ends of the two liquid inlet diversion pipelines 161 are communicated and are both communicated with the sample inlet 110. The outlet ends of the two inlet flow-dividing pipes 161 are not communicated.

The second-stage liquid inlet branch pipe 163 includes four liquid inlet branch pipes 161 arranged in parallel, and may be divided into two groups. One set of these inlet liquid diversion conduits includes two inlet liquid diversion conduits 161 located at the upper side of the inlet liquid diversion conduit in fig. 19, and the inlet ends of these two inlet liquid diversion conduits 161 are communicated with the outlet end of one of the two inlet liquid diversion conduits 161 of the first stage inlet liquid diversion conduit 162. The other set comprises two inlet liquid diversion pipes 161 which are positioned at the lower part of the liquid inlet pipe 161 in the figure 19, and the inlet ends of the two inlet liquid diversion pipes 161 are communicated with the outlet end of the other inlet liquid diversion pipe 161 of the first stage inlet liquid diversion pipe 162. The outlet ends of the four inlet flow-dividing pipes 161 of the second stage are respectively communicated with one inlet groove 120.

Each of the feed split pipes 161 located in the upper stage communicates with two of the feed split pipes 161 located in the lower stage. A plane perpendicular to the flow guiding direction of the liquid inlet branch pipe 161 is taken as a cross section, and the cross section area of the liquid inlet branch pipe 161 at the next stage is half of the cross section area of the liquid inlet branch pipe 161 at the previous stage. So set up, can improve the homogeneity that the sample solution flows in the feed liquor slot 120 for pressure is balanced in each feed liquor slot 120, avoids pressure too big in a certain feed liquor slot 120, leads to chip body 100 to destroy and can not normally work.

In another possible example, referring to FIG. 20, the inlet connection conduit 160 comprises a three stage inlet bleed conduit. The first-stage liquid inlet shunting pipeline close to the sample inlet 110 comprises two liquid inlet shunting pipelines 161 which are arranged in parallel, and the inlet ends of the two liquid inlet shunting pipelines 161 are communicated and are both communicated with the sample inlet 110. The outlet ends of the two inlet flow-dividing pipes 161 are not communicated.

The middle one-stage liquid inlet branch pipe includes four liquid inlet branch pipes 161 arranged in parallel, and the inlet ends of the two liquid inlet branch pipes 161 located above the liquid inlet branch pipe shown in fig. 20 are communicated with the outlet end of one liquid inlet branch pipe 161 close to the sample inlet 110. The inlet ends of two inlet branch pipes 161 located at the lower side in fig. 20 are communicated with each other, and the outlet ends of the other inlet branch pipe 161 close to the sample inlet 110 are communicated with each other, and the outlet ends of the four inlet branch pipes 161 are not communicated with each other.

The first inlet flow dividing line adjacent to the inlet channel 120 comprises eight inlet flow dividing lines 161 arranged in parallel. The eight liquid inlet branch pipes 161 are grouped into four groups by adjacent two, and the inlet ends of the two liquid inlet branch pipes 161 in each group are communicated with the outlet end of each liquid inlet branch pipe 161 in the middle stage. The outlet ends of the eight inlet flow dividing pipes 161 are respectively communicated with the inlet grooves 120. The plane perpendicular to the flow guiding direction of the liquid inlet distribution pipeline 161 is taken as a cross section, and in the adjacent two stages of liquid inlet distribution pipelines, the cross section of the liquid inlet distribution pipeline positioned at the next stage is half of the cross section of the liquid inlet distribution pipeline positioned at the previous stage.

A liquid outlet connecting pipe 170 may be further disposed between the liquid outlet groove 140 and the sample outlet 150. The outlet connecting pipe 170 is used for converging, and the sample solution in the plurality of outlet grooves 140 is converged to the sample outlet 150. Referring to fig. 20, the outlet connection pipe 170 includes at least two stages of outlet diversion pipes arranged in sequence along the direction from the sample outlet 150 to the outlet groove 140. Each level of liquid outlet diversion pipeline comprises at least two liquid outlet diversion pipelines 171 which are arranged in parallel, and each liquid outlet diversion pipeline 171 positioned in the upper level is communicated with at least two liquid outlet diversion pipelines 171 positioned in the lower level. The first-stage liquid outlet diversion pipeline close to the sample outlet 150 is communicated with the sample outlet 150, and the first-stage liquid outlet diversion pipeline close to the liquid outlet groove 140 is respectively communicated with the liquid outlet groove 140. The specific structure of the liquid outlet connecting pipe 170 can refer to fig. 19 and 20 and the structure of the liquid inlet connecting pipe 160, and the detailed description is omitted.

For example, only the liquid inlet connection pipe 160, only the liquid outlet connection pipe 170, or both the liquid inlet connection pipe 160 and the liquid outlet connection pipe 170 may be disposed in the chip body 100. The number of stages of the liquid inlet connecting pipe 160 and the liquid outlet connecting pipe 170 may be the same or different. For example, the inlet connection pipe 160 includes two stages, and the outlet connection pipe 170 includes four stages. When the number of stages of the inlet connection pipe 160 and the outlet connection pipe 170 in the chip body 100 is the same, the flow uniformity of the sample solution is better.

In the cell screening chip provided by the embodiment of the present disclosure, the chip body 100 includes a liquid inlet channel 120, a liquid outlet channel 140 and a screening array 130, and the screening array 130 is located between the liquid inlet channel 120 and the liquid outlet channel 140. The screening array 130 comprises a plurality of screening units, each screening unit comprises a containing cavity 131 and a screening channel 132, and the liquid inlet groove 120, the containing cavity 131, the screening channel 132 and the liquid outlet groove 140 are sequentially communicated. The sample solution containing the target cells flows into the containing cavity 131 and the screening channel 132 from the liquid inlet channel 120 in sequence, and flows out from the liquid outlet channel 140. Since the width of the accommodating chamber 131 is larger than the diameter of the target cell and the width of the screening channel 132 is smaller than the diameter of the target cell, the target cell cannot be trapped in the accommodating chamber 131 through the screening channel 132. Non-target cells having a diameter smaller than the width of the screening channel 132 flow out of the screening channel 132 to achieve separation of target cells from non-target cells. Meanwhile, in each screening unit, an included angle between the diversion direction of the accommodating cavity 131 and the diversion direction of the liquid inlet groove 120 is an acute angle, so that the sample solution in the liquid inlet groove 120 stably flows into the accommodating cavity 131, vortexes and backflow in the sample solution are reduced, the target cells are prevented from being impacted, the deformation of the target cells is prevented from extruding the screening channel 132, the rejection rate of the target cells is improved, and the capture rate of the target cells is improved.

Example two

Referring to fig. 21, embodiments of the present disclosure provide a cell screening system for separating and identifying target cells. The cell screening system comprises the cell screening chip 10, a sample pump 20 and a waste liquid collecting device 70. The sample inlet of the cell screening chip 10 is connected to the sample pump 20, and the sample outlet is connected to the waste liquid collecting device 70. The sample pump 20 is used for pumping the sample solution, the cell screening chip 10 is used for capturing the target cells, so as to separate the target cells from the sample solution, and the waste liquid collecting device 70 is used for collecting the waste liquid flowing out from the cell screening chip 10.

The sample 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 diluent may be Phosphate Buffer Saline (PBS). The blood sample and the diluent enter the cell screening chip 10 through the structure of double injection ports to be mixed, so that the time required by detection is shortened. In another possible example, the sample solution pump 22 pumps a diluted blood sample.

The sample pump 20 may further include one or more of a surface treatment liquid pump 21, a buffer liquid pump 23, a stationary liquid pump 24, and a dye liquid pump 25. For example, as shown in FIG. 21, the sample pump 20 includes a processing liquid pump 21, a sample solution pump 22, a buffer liquid pump 23, a stationary liquid pump 24, and a staining liquid pump 25, and pumps different liquids to the cell screening chip 10.

When the sample pump 20 includes a plurality of pumps, a reversing valve 30 is disposed between the sample pump 20 and the cell screening chip 10. That is, the output end of the sample pump 20 is connected with one end of the reversing valve 30, the other end of the reversing valve 30 is connected with the sample inlet of the cell screening chip 10, and the liquid in each pump enters the cell screening chip 10 according to a certain sequence through the reversing valve 30.

For example, the surface treatment solution may be polyvinylpyrrolidone (PVP) for reducing the flow resistance of the sample solution. The buffer may be the same as the diluent, and may be PBS. The fixative solution may be a solution containing 4% Paraformaldehyde (PFA) for the purpose of typing target cells. Specifically, the fixing liquid can reduce the elasticity of cells, the cells are not easy to deform after the action of the fixing liquid, and various structures in the cells are fixed, so that the cells are shaped. The staining solution may be a fluorescent stain for staining the target cells for identification, e.g., when the target cells are circulating tumor cells, the fluorescent stain may comprise CD with a fluorescein₄₅4',6-diamidino-2-phenylindole (DAPI) and Epithelial Cell Adhesion Molecule (EpCAM) with another fluorescein, in which CD is the ligand of the formula₄₅For labeling leukocytes, EpCAM for labeling circulating tumor cells, DAPI for labeling nuclei, and further identification of target cells by staining different cells.

With continued reference to fig. 21, 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 for illuminating the cell screening chip 10 when the image capturing device 50 is in operation, and the light source 40 may be an LED lamp, or an incandescent lamp or a neon lamp, etc., and provides background light. The light source 40 and the image capturing device 50 may be located on the same side of the cell screening chip 10, or may be located on both sides of the cell screening chip 10. For example, in the present embodiment, the light source 40 is located at the lower side of the cell screening chip 10, and the image capturing device 50 is located at the upper side of the cell screening chip 10.

The image acquisition device 50 is in signal connection with the data processing device 60, and is used for acquiring the image of the cell screening chip 10 and transmitting the image to the data processing device 60. The image capturing Device 50 may be a Charge-coupled Device (CCD), and the data processing Device 60 may be a computer for identifying the number of target cells.

With continued reference to fig. 21, the cell screening system may further include a stage 80, the stage 80 being used to place the cell screening chip 10. The carrier 80 may be a conveyor belt to move the cell screening chip 10 relative to the image capturing device 50 at a certain speed, so as to ensure that the image capturing device 50 can capture the image of the whole cell screening chip 10.

In the embodiment of the present disclosure, the cell screening system includes a sample pump 20, a waste liquid collecting device 70, and the cell screening chip 10. The sample inlet of the cell screening chip 10 is connected to the sample pump 20, and the sample outlet of the cell screening chip 10 is connected to the waste liquid collecting device 70. The cell screening system comprises the cell screening chip, so that the cell screening system has the advantage of high capture rate of target cells, and specific effects are referred to above and are not repeated herein.

EXAMPLE III

Referring to fig. 22, an embodiment of the present disclosure provides a cell screening method, to which the above cell screening system is applied, for separating and identifying target cells, the cell screening method including:

s101, injecting surface treatment liquid and diluent into the cell screening chip 10 in sequence for pretreatment.

Surface treatment liquid is pumped into the cell screening chip 10 through the sample pump 20 and flows out of the waste liquid collecting device 70, and the functional area of the cell screening chip 10 for capturing target cells is subjected to surface treatment, wherein the surface treatment liquid can be PVP.

And pumping the diluent to the cell screening chip 10 through the sample pump 20, flushing the surface treatment solution, filling the cell screening chip 10 with the surface treatment solution, and exhausting the bubbles, wherein the diluent can be PBS.

S102, injecting a sample solution containing the target cells into the cell screening chip 10, and capturing the target cells by the cell screening chip 10.

In the embodiment of the present disclosure, the diluted blood sample may be pumped into the cell screening chip 10 by the sample pump 20, that is, the mixed sample solution may be pumped into the cell screening chip 10, or the blood sample and the diluent may be respectively pumped into the cell screening chip 10 by the sample pump 20 to be mixed to form the sample solution.

The cell sorting chip 10 is used to separate target cells from a sample solution, non-target cells having a smaller size flow out of the cell sorting chip 10, and target cells having a larger size are captured and retained by the cell sorting 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 capture rate of target cells is high.

S103, sequentially injecting a diluent, a fixing solution, a diluent, a staining solution and a diluent into the cell screening chip 10, and typing and staining the target cells.

In the embodiment of the present disclosure, the sample pump 20 pumps the diluent into the cell screening chip 10 to clean the cell screening chip 10; pumping fixing solution into the cell screening chip 10 by a sample pump 20 to shape the target cell, wherein the fixing solution can be PFA; pumping the diluent into the cell screening chip 10 by the sample pump 20, cleaning the cell screening chip 10 again, and cleaning the fixing solution; the sample pump 20 pumps the staining solution into the cell screening chip 10 to label the target cells, and further distinguish the cell types in the cell screening chip 10, wherein the staining solution can be a fluorescent staining agent.

For example, the sample solution contains circulating tumor cells, red blood cells, platelets, and white blood cells, the diameter of the circulating tumor cells is about 10-20 μm, the diameter of the red blood cells is about 6-9 μm, the diameter of the platelets is about 1-4 μm, and the diameter of the white blood cells is about 7-20 μm. When the target cells are circulating tumor cells, the cell selection chip 10 captures the circulating tumor cells and a part of leukocytes. In order to distinguish the two cells, the two cells can be labeled with different fluorescent colors by a staining solution so as to identify the circulating tumor cells.

After dyeing, the sample pump 20 pumps the diluent into the cell screening chip 10, the cell screening chip 10 is cleaned again, and the dyeing solution is washed clean, so as to collect the fluorescent image of the cell screening chip 10 subsequently.

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 cell.

In the embodiment of the present disclosure, when the image acquisition device 50 acquires the fluorescence 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 light is supplemented for the image acquisition device 50, so that the image acquisition device 50 can acquire a clearer image.

The image acquisition device 50 is in signal connection with the data processing device 60 and transmits the image to the data processing device 60, and the data processing device 60 identifies the number of target cells.

In the embodiment of the present disclosure, the cell screening chip 10 is injected with the sample solution after the surface treatment solution and the diluent are sequentially injected for pretreatment, 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 cell screening system, the capture rate of the target cells can be improved, and details are not repeated herein. Meanwhile, the target cells are shaped by using the fixing solution, and are subjected to staining identification by using the staining solution so as to be distinguished from non-target cells, so that the target cells are conveniently identified.

The embodiments or implementation modes in the present specification are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments may be referred to each other.

It will be understood by those skilled in the art that in the present disclosure, the terms "longitudinal," "lateral," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in an orientation or positional relationship indicated in the drawings for ease of description and simplicity of description, and do not indicate or imply that the system or element being referred to must be in a particular orientation, constructed and operated in a particular orientation, and therefore, the above terms should not be construed as limiting the present disclosure.

In the description of the present specification, references to "one embodiment", "some embodiments", "an illustrative embodiment", "an example", "a specific example", or "some examples", etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present disclosure. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

Finally, it should be noted that: the above embodiments are only used for illustrating the technical solutions of the present disclosure, and not for limiting the same; while the present disclosure has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art will understand that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present disclosure.

Claims

1. A cell screening chip is characterized by comprising a chip body, wherein the chip body is provided with a liquid inlet groove and a liquid outlet groove, the tail end of the liquid inlet groove is closed along the flow guide direction of the liquid inlet groove, and a screening array is formed between the side walls of the liquid inlet groove and the liquid outlet groove, which are close to each other;

the screening array comprises a plurality of screening units, each screening unit comprises an accommodating cavity and a screening channel, the inlet end of the accommodating cavity is communicated with the liquid inlet groove, the outlet end of the accommodating cavity 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 larger than the diameter of the target cell, and the width of the screening channel is smaller than the diameter of the target cell;

in each screening unit, an included angle between the flow guide direction of the accommodating cavity and the flow guide direction of the liquid inlet groove is an acute angle.

2. The cell screening chip of claim 1, wherein in each screening unit, the flow direction of the accommodating cavity coincides with or is parallel to the flow direction of the screening channel.

3. The cell screening chip of claim 1, wherein the screening channel is a flow guide groove or a flow guide hole formed in the chip body.

4. The cell screening chip according to claim 3, wherein the outlet end of the screening channel is formed with a buffer groove, and the flow direction of the buffer groove coincides with the flow direction of the screening channel; the orthographic projection of the buffer groove on a plane perpendicular to the flow guide direction of the screening channel is larger than the orthographic projection of the screening channel on the plane.

5. The cell screening chip according to claim 1, wherein the screening array is formed in a chip body between the inlet channel and the outlet channel.

6. The cell screening chip of claim 5, wherein the width of the receiving cavity at the middle of the screening array is greater than the width of the receiving cavity at the end of the screening array.

7. The cell screening chip of claim 5, wherein the width of the receiving cavity is gradually reduced along a direction from the middle to the end of the screening array.

8. The cell screening chip of claim 1, wherein the chip body is further provided with a sample inlet and a sample outlet, the sample inlet is communicated with the liquid inlet groove, and the sample outlet is communicated with the liquid outlet groove.

9. The cell screening chip of claim 8, wherein the sample inlet comprises a first sample inlet and a second sample inlet respectively communicating with the liquid inlet groove.

10. The cell screening chip of claim 8, wherein the chip body is further provided with a liquid inlet connecting pipeline between the liquid inlet groove and the sample inlet;

the liquid inlet connecting pipeline comprises at least two stages of liquid inlet shunting pipelines which are sequentially arranged along the direction from the sample inlet to the liquid inlet groove, each stage of liquid inlet shunting pipeline comprises at least two liquid inlet shunting pipelines which are distributed in parallel, and each liquid inlet shunting pipeline in the previous stage is communicated with at least two liquid inlet shunting pipelines in the next stage;

and in the feed liquor reposition of redundant personnel pipeline at each level, be closest the one-level of introduction port feed liquor reposition of redundant personnel pipeline with introduction port intercommunication, be closest the one-level of feed liquor slot feed liquor reposition of redundant personnel pipeline respectively with feed liquor slot intercommunication.

11. The cell screening chip according to claim 8 or 10, wherein the chip body is further provided with a liquid outlet connecting pipe between the liquid outlet groove and the sample outlet;

the liquid outlet connecting pipeline comprises at least two stages of liquid outlet distribution pipelines which are sequentially arranged along the direction from the sample outlet to the liquid outlet groove, and each stage of liquid outlet distribution pipeline comprises at least two liquid outlet distribution pipelines which are distributed in parallel; each liquid outlet distribution pipeline in the upper stage is communicated with at least two liquid outlet distribution pipelines in the lower stage;

and in each level of the liquid outlet diversion pipelines, the liquid outlet diversion pipeline is closest to the first level of the sample outlet and communicated with the sample outlet, and the liquid outlet diversion pipeline is closest to the first level of the liquid outlet groove and communicated with the liquid outlet groove respectively.

12. The cell screening chip according to claim 1, wherein the chip body is provided with one inlet channel, and at least one of both sides of the inlet channel is provided with one outlet channel.

13. The cell screening chip according to any one of claims 1, 2, 5 to 10 and 12, wherein the liquid inlet channel and the liquid outlet channel are serpentine, the liquid inlet channel comprises at least two linear channels parallel to each other, and in each two adjacent linear channels, the outlet of one of the linear channels is communicated with the inlet of the other linear channel through a first arc-shaped channel;

the screening array is arranged on the side face of the linear channel and/or the side face of the first arc-shaped channel.

14. The cell screening chip according to any one of claims 1, 2, 5 to 10 and 12, wherein the liquid inlet groove and the liquid outlet groove are both wavy lines, the liquid inlet groove comprises at least two second arc-shaped channels which are communicated in sequence, and the screening array is arranged on the side surface of each second arc-shaped channel.

15. The cell screening chip of claim 14, wherein the wavy line is formed by connecting semi-circles in sequence, or the wavy line is one of a sine curve and a cosine curve.

16. The cell screening chip according to any one of claims 1, 2, 5 to 10 and 12, wherein the inlet channel is linear, and the width of the inlet channel gradually increases along the direction from the middle of the inlet channel to the end of the inlet channel.

17. The cell screening chip according to any one of claims 1, 2, 5 to 10 and 12, wherein the liquid inlet groove and the liquid outlet groove are both spiral-shaped, and the liquid inlet groove and the liquid outlet groove are engaged, and the liquid outlet groove is located on a side of the liquid inlet groove away from a center of the spiral.

18. The cell screening chip of claim 17, wherein the inlet channel is connected to a first inlet and a second inlet respectively, the first inlet is disposed on a side of the inlet channel close to the screening array, and the second inlet is disposed on a side of the inlet channel away from the screening array.

19. A cell screening system, comprising a sample pump, a waste liquid collecting device, and the cell screening chip of any one of claims 1 to 18, wherein a sample inlet of the cell screening chip is connected to the sample pump, and a sample outlet of the cell screening chip is connected to the waste liquid collecting device.

20. The cell screening system of claim 19, wherein the sample pump comprises a sample solution pump and one or more of a surface treatment solution pump, a buffer solution pump, a stationary solution pump, and a staining solution pump;

the output end of the sample injection pump is connected with one end of a reversing valve, and the other end of the reversing valve is connected with the sample injection port.

21. The cell screening system of claim 19, further comprising a light source, an image acquisition device, and a data processing device;

the light source is used for irradiating the cell screening chip when the image acquisition device works;

the image acquisition device is in signal connection with the data processing 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 according to the images.

22. A cell screening method using the cell screening system according to any one of claims 19 to 21, the cell screening method comprising:

sequentially injecting surface treatment liquid and buffer liquid into the cell screening chip for pretreatment;

injecting a sample solution containing target cells into the cell screening chip, which captures the target cells;

sequentially injecting a buffer solution, a fixing solution, a buffer solution, a staining solution and a buffer solution into the cell screening chip, and typing and staining the target cells;

the image acquisition device acquires an image of the cell screening chip and transmits the image to the data processing device, and the data processing device identifies the target cell.