WO2022088966A1

WO2022088966A1 - Cell screening chip, and cell screening system and method

Info

Publication number: WO2022088966A1
Application number: PCT/CN2021/116108
Authority: WO
Inventors: 杨旸; 庄紫云; 姜奥
Original assignee: 上海荧辉医疗器械有限公司
Priority date: 2020-10-29
Filing date: 2021-09-02
Publication date: 2022-05-05
Also published as: CN112175792A

Abstract

The present disclosure relates to the technical field of biological detection, and provides a cell screening chip, and a cell screening system and method, used for solving the technical problem of a low capture rate of target cells. A chip body of the cell screening chip is provided with a liquid inlet groove, a tail end of the liquid inlet groove is closed in a flow guide direction of the liquid inlet groove, the two sides of the liquid inlet groove are each provided with a liquid outlet groove, and a screening array is arranged between the liquid inlet groove and each liquid outlet groove. Each screening unit of each screening array comprises an accommodating cavity and a screening channel, and the liquid inlet groove, the accommodating cavity, the screening channel, and the corresponding liquid outlet groove are sequentially communicated. The diameter of target cells is greater than the width of the screening channels and less than the width of the accommodating cavities. Inlet ends of the accommodating cavities in one of the two screening arrays and inlet ends of the accommodating cavities in the other screening array are arranged in a staggered manner, so that the liquid inlet groove realizes staggered diversion; the target cells flow through one accommodating cavity each time and are affected by the lateral flow velocity in one direction, thereby reducing the probability that the target cells avoid the accommodating cavities, and increasing the capture rate.

Description

Cell screening chip, cell screening system and method thereof

This application claims the priority of the Chinese patent application with the application number 202011186434.7 and the application title "Cell Screening Chip, Cell Screening System and Method" filed with the China Patent Office on October 29, 2020, the entire contents of which are incorporated by reference in in this application.

technical field

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

Background technique

In the field of biological detection, it is often necessary to separate the target cells for further observation or testing of the target cells, for example, the separation and detection of circulating tumor cells. Circulating Tumor Cells (CTCs) are tumor cells that originate from primary tumors and enter the circulatory system. Such cells can develop into metastases through the circulatory system, and then multiply to form secondary tumors. , the timely isolation and detection of such cells is of great significance for monitoring the treatment and recurrence of tumors.

For the screening of target cells, they can be separated from the sample solution based on the size of the target cells. Usually, a microfilter with a channel size smaller than the diameter of the target cells is designed to capture the target cells with a larger size, and other cells with a smaller size flow out with the buffer. to isolate the target cells. However, target cells with this method are prone to rupture as the buffer flows through the microfilter, resulting in a low capture rate of target cells.

SUMMARY OF THE 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, which are used to improve the capture rate of target cells.

In order to achieve the above purpose, 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, the chip body is provided with a liquid inlet groove, and along the flow direction of the liquid inlet groove, the end of the liquid inlet groove is closed, two sides of the liquid inlet groove are respectively provided with a liquid outlet groove, and a screening array is formed between the liquid inlet groove and each of the liquid outlet grooves; A plurality of screening units are arranged in the direction of the liquid inlet groove, each screening unit includes an accommodating cavity and a screening channel, the inlet end of the accommodating cavity is communicated with the liquid feeding groove, and the accommodating cavity is in communication with the liquid feeding groove. The outlet end of the screening channel 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; and 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; in two of the screening arrays, the inlet end of the accommodating cavity in one of the screening arrays and the inlet of the accommodating cavity in the other of the screening arrays end misalignment settings.

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

In the cell screening chip provided by the embodiment of the present disclosure, the chip body includes a liquid inlet groove, along the extending direction of the liquid inlet communication, the end of the liquid inlet groove is closed, and the liquid inlet groove is provided with a liquid outlet groove on both sides. A screening array is arranged between the liquid groove and each liquid outlet groove. The screening array includes a plurality of screening units arranged along the diversion direction of the liquid inlet groove, each screening unit includes a accommodating cavity and a screening channel, and the liquid inlet groove, the accommodating cavity, the screening channel and the liquid outlet groove are connected in sequence, and contain The sample solution of the target cells flows into the accommodating cavity and the screening channel in sequence from the liquid inlet groove, and flows out from the liquid outlet groove. Since 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, the target cells cannot pass through the screening channel and are trapped in the accommodating cavity. Flow out to achieve separation of target cells from non-target cells. At the same time, since the inlet ends of the accommodating cavities located on both sides of the liquid inlet groove are dislocated, compared with the symmetrical distribution of the accommodating cavities on both sides of the liquid inlet groove, in the embodiment of the present disclosure, the two sides of the liquid inlet groove The lateral accommodating cavities are staggered, so that the accommodating cavities located on both sides of the liquid inlet groove are staggered and shunted, and the target cells only flow through one accommodating cavity at a time, which is affected by the lateral flow rate in one direction, reducing the missed capacity of the target cells. The probability of placing the cavity increases the retention rate of target cells, thereby improving the capture rate of target cells.

In a second aspect, an embodiment of the present disclosure further provides a cell screening system, including a sample injection pump, a waste liquid collection device, and the above-mentioned cell screening chip, wherein an injection port of the cell screening chip is connected to the sample injection pump , the sample outlet of the cell screening chip is connected to the waste liquid collection device.

The cell screening system provided by the embodiments of the present disclosure includes the above-mentioned cell screening chip, and thus also has the advantage of high capture rate of target cells. The specific effects can be referred to above, and will not be repeated here.

In a third aspect, an embodiment of the present disclosure also provides a cell screening method using the above cell screening system, the cell screening method comprising: sequentially injecting a surface treatment solution and a buffer solution into a cell screening chip for pretreatment; The sample solution is injected into the cell screening chip, and the cell screening chip captures the target cells; the cell screening chip is sequentially injected with a buffer, a fixative, a buffer, a staining solution and a buffer, and the target cells are shaped and stained; An image acquisition device acquires an image of the cell screening chip, and transmits the image to a data processing device, and the data processing device identifies the target cell.

The cell screening method provided by the embodiments of the present 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 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, it can improve the The capture rate of target cells will not be repeated here. At the same time, the fixative solution is used to finalize the target cells, and the staining solution is used to stain and identify the target cells to distinguish them from non-target cells, which is convenient for the identification of the target cells.

Description of drawings

FIG. 1 is a schematic structural diagram of a cell screening chip in an embodiment of the disclosure;

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

Fig. 3 is the partial enlarged view of A place in Fig. 2;

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

5 is a schematic structural diagram of a screening unit in an embodiment of the present disclosure;

Fig. 6 is a partial enlarged view at B in Fig. 5;

7 is another schematic structural diagram of a screening unit in an embodiment of the present disclosure;

Fig. 8 is a partial enlarged view at C in Fig. 7;

FIG. 9 is a first structural schematic diagram of a liquid inlet groove in an embodiment of the present disclosure;

Fig. 10 is the first partial enlarged view at D in Fig. 9;

Fig. 11 is the second kind of partial enlarged view at D place in Fig. 9;

Fig. 12 is the third partial enlarged view at D in Fig. 9;

13 is a schematic diagram of the second structure of the liquid inlet groove in the embodiment of the present disclosure;

14 is a schematic diagram of a third structure of the liquid inlet groove in the embodiment of the present disclosure;

Fig. 15 is a partial enlarged view at E in Fig. 14;

16 is a schematic structural diagram of a dual injection port in an embodiment of the present disclosure;

17 is a schematic structural diagram of a liquid inlet connection channel in an embodiment of the present disclosure;

18 is another schematic structural diagram of the liquid inlet connection channel in the embodiment of the disclosure;

19 is a schematic structural diagram of a cell screening system in an embodiment of the disclosure;

FIG. 20 is a schematic flowchart of a cell screening method in an embodiment of the disclosure.

Description of reference numbers:

10. Cell screening chip; 100. Chip body;

110. Injection port; 111. The first injection port;

112. The second injection port; 120. The liquid inlet groove;

121. Straight channel; 122. The first arc channel;

123. Arc-shaped deflector; 124. Second arc-shaped channel;

130, screening array; 131, accommodating cavity;

132, screening channel; 133, buffer tank;

140. Liquid outlet groove; 150. Sample outlet;

160. Inlet connection pipeline; 161. Inlet shunt pipeline;

162. The first-stage liquid inlet shunt pipeline; 163. The second-stage liquid inlet shunt pipeline;

170. Outlet connection pipeline; 171. Outlet shunt pipeline;

180. Cover; 20. Sampling pump;

21. Surface treatment liquid pump; 22. Sample solution pump;

23. Buffer pump; 24. Fixed liquid pump;

25. Dyeing liquid pump; 30. Reversing valve;

40. Light source; 50. Image acquisition device;

60. Data processing device; 70. Waste liquid collection device;

80. Carrier; L1, the width of the accommodating cavity;

L2, the width of the screening channel.

Detailed ways

In the related art, a cell screening chip includes a chip body and a cover, the chip body and the cover are sealed, and there is a channel between the chip body and the cover for screening target cells, however, the cell screening chip has a capture rate of target cells. Low technical issues.

In view of the above technical problems, the cell screening chip in the embodiment of the present disclosure includes a chip body, the chip body is provided with a liquid inlet groove, and the end of the liquid inlet groove is closed along the flow direction of the liquid inlet groove. Two sides of the liquid inlet groove are respectively provided with a liquid outlet groove, and a plurality of screening units are arranged between the liquid inlet groove and the liquid outlet groove. Each screening unit includes a accommodating cavity and a screening channel. The accommodating cavity, the screening channel and the liquid outlet groove are connected in sequence, 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. The inlet ends of the accommodating cavities located on both sides of the liquid inlet groove are staggered, so that the accommodating cavities are distributed in a staggered manner, and then the flow is staggered. The target cells flow through one accommodating cavity at a time, and are affected by the lateral flow rate in one direction, reducing the The probability of missing the accommodating cavity is reduced, and the capture rate of target cells is improved.

In order to make the above objects, features and advantages of the embodiments of the present disclosure more obvious and easy to understand, the technical solutions in the embodiments of the present disclosure will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present disclosure. Obviously, the described embodiments are only some, but not all, embodiments of the present disclosure. Based on the embodiments in the present disclosure, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present disclosure.

Example 1

In order to separate the target cells in the sample solution for subsequent observation or inspection, an embodiment of the present disclosure provides a cell screening chip. The material of the cell screening chip can be a transparent material 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 cover 180 . The cover 180 covers the chip body and is used to seal the chip body 100 to prevent the sample solution from flowing out of the cell screening chip.

2, the end face of the chip body 100 is formed with a sample inlet 110, a liquid inlet groove 120, a screening array 130 (shown by the dotted line in Figure 2), a liquid outlet groove 140 and a sample outlet 150. Specifically, the chip The end surface of the main body 100 is the upper surface of the chip main body 100 . Correspondingly, the cover 180 covers the upper surface of the chip main body 100 .

Along the diversion direction of the liquid inlet groove 120 , the end of the liquid inlet groove 120 is closed. As shown in FIG. 2 , the left end of the liquid inlet groove 120 is connected to the sample inlet 110 , and the right end of the liquid inlet groove 120 is blocked. Two sides of the liquid inlet groove 120 are respectively provided with a liquid outlet groove 140 , and a screening array 130 is arranged between the liquid inlet groove 120 and the liquid outlet groove 140 on each side, and is communicated by the screening array 130 . That is, the liquid inlet groove 120 communicates with the two liquid outlet grooves 140 , and a screening array 130 is disposed between the liquid inlet groove 120 and each liquid outlet groove 140 .

One end of the two liquid outlet grooves 140 is communicated with the sample outlet 150 , and the other end is blocked, that is, the two liquid outlet grooves 140 converge at the sample outlet 150 . As shown in FIG. 2 , the left end of the liquid outlet groove 140 is blocked, and the right end of the liquid outlet groove 140 is communicated with the sample outlet 150 . The sample solution flows into the sample inlet 110 and flows through the liquid inlet groove 120, the screening array 130 and the liquid outlet groove 140 in sequence to the sample outlet 150. The screening array 130 is used to intercept and capture the target cells.

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 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 extending direction of the liquid inlet groove 120 , and the liquid inlet groove 120 The end of the liquid inlet groove 120 refers to the starting or ending face of the extending direction of the liquid inlet groove 120 . It can be understood that the side walls of the liquid inlet groove 120 are connected to both ends of the liquid inlet groove 120 . The structure of the liquid outlet groove 140 is similar to that of the liquid inlet groove 120 , and details are not described herein again.

Taking the structure shown in FIG. 1 as an example, the left and right sides of the liquid inlet groove 120 are the ends of the liquid inlet groove 120 , and the front and rear vertical surfaces of the liquid inlet groove 120 are the side walls of the liquid inlet groove 120 . 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 .

Taking the orientation shown in FIG. 2 as an example, the two sides of the liquid inlet groove 120 are provided with a liquid outlet groove 140 respectively, which means that the upper and lower sides of the liquid inlet groove 120 are provided with a liquid outlet groove 140 . The liquid inlet groove 120 and the liquid outlet groove 140 are arranged side by side, and 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 is communicated by the screening array 130 . As shown in FIG. 2 , a screening array 130 is provided between the upper side wall of the liquid inlet groove 120 and the lower side wall of the liquid outlet groove 140 located on the upper side of the liquid inlet groove 120 . A screening array 130 is disposed between the side wall and the upper side wall of the liquid outlet groove 140 located on the lower side of the liquid inlet groove 120 .

The extending direction of the liquid inlet groove 120 is parallel to the extending direction of the liquid outlet groove 140, that is, the flow direction of the liquid inlet groove 120 is parallel to the flow direction of the liquid outlet groove 140, so that the flow uniformity of the sample solution is relatively high. Well, the diversion direction is related to the channel shape.

In the embodiments of the present disclosure and the following embodiments, the extending direction of the liquid inlet groove 120 is also the direction of the flow of the liquid inlet groove 120 , and the extending direction of the liquid outlet groove 140 is also the guiding direction of the liquid outlet groove 140 . The flow direction is not repeated here.

Continuing to refer to FIG. 2 , the screening array 130 includes a plurality of screening units, and the plurality of screening units are arranged along the diversion direction of the liquid inlet groove 120 for intercepting and capturing target cells. The number of screening units in the screening array 130 located on both sides of the liquid inlet groove 120 may be the same or different. In this embodiment, the screening arrays 130 located on both sides of the liquid inlet groove 120 include seven screening units.

Referring to FIG. 3 , each screening unit includes an accommodating cavity 131 and a screening channel 132 communicating with the accommodating cavity 131 . The liquid inlet groove 120 , the accommodating cavity 131 , the screening channel 132 and the liquid outlet groove 140 are connected in sequence. That is, the liquid inlet groove 120 communicates with the inlet end of the accommodating cavity 131 , the outlet end of the accommodating cavity 131 communicates with the inlet end of the screening channel 132 , and the outlet end of the screening channel 132 communicates with the liquid outlet groove 140 . The liquid inlet groove 120 , the accommodating cavity 131 , the screening channel 132 and the liquid outlet groove 140 form a channel path for the sample solution to flow, so that the sample solution flows through the cell screening chip through these channel paths.

Continuing to refer to FIG. 2 , in the two screening arrays 130 located on opposite sides of the liquid inlet groove 120 , the inlet end of the accommodating cavity 131 in one of the screening arrays 130 and the inlet end of the accommodating cavity 131 in the other screening array The staggered arrangement makes the flow in the liquid inlet groove 120 staggered. The arrow in FIG. 2 shows the shunt path of the liquid inlet groove 120 . Compared with the symmetrical arrangement of the accommodating cavities 131 on both sides, the flow distribution length of the liquid inlet groove 120 in the embodiment of the present disclosure is increased, that is, the liquid inlet groove 120 The whole process is diverted, which improves the diversion efficiency. The target cells flow through one accommodating cavity 131 at a time, which reduces the probability of missing the accommodating cavity 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 generally laminar, the flow rate of the sample solution close to the centerline of the liquid inlet groove 120 is large, and the flow rate of the sample solution far from the centerline of the liquid inlet groove 120 is small, The accommodating cavity 131 is laterally divided to provide a lateral flow rate. Since the target cells are affected by the lateral flow rate of one accommodating cavity 131 at a time, the target cells flow into the accommodating cavity 131, which improves the capture rate of the target cells.

The accommodating cavities 131 in the screening array 130 located on each side of the liquid inlet groove 120 can be uniformly arranged along the diversion direction of the liquid feeding groove 120 , that is, the accommodating cavities 131 on each side have the same value. spacing. Each accommodating cavity 131 on one side of the liquid inlet groove 120 has a first spacing, and each accommodating cavity 131 on the other side of the liquid feeding groove 120 has a second spacing, the first spacing and the second spacing can be equal, or may not be equal.

In some possible examples, in two screening arrays 130 located on opposite sides of the liquid inlet groove 120, the inlet end of the accommodating cavity 131 in one screening array 130 and the inlet of the accommodating cavity 131 in the other screening array The ends can be completely staggered, that is, the orthographic projections of the inlet ends of the accommodating chambers 131 on both sides of the liquid inlet groove 120 in the direction of the flow of the liquid inlet groove 120 are completely non-overlapping. At this time, the orthographic projections of the accommodating cavities 131 on both sides of the liquid inlet groove 120 on the flow direction of the liquid inlet groove 120 may be continuous or may have intervals.

In some other possible examples, 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 screening array 130 is the same as the inlet end of the accommodating cavity 131 in the other screening array 130 . The inlet ends may be partially staggered, that is, the orthographic projections of the inlet ends of the accommodating chambers 131 on both sides in the direction of the flow of the liquid inlet groove 120 partially overlap.

In the above-mentioned embodiment, the cover 180 may be located above the chip body 100 , or may be located under the chip body 100 . Specifically, the cover 180 is covered on the chip body 100, and the bottom surface of the cover 180 is abutted with the upper surface of the chip body 100, so as to prevent the target cells from overflowing into the chip body 100 from the gap between the two surfaces to be used for the target The cell screening area improves the retention rate of target cells, thereby improving the detection accuracy of the cell screening chip.

The cell screening chip can be a microfluidic chip, and its material can be transparent polymethyl methacrylate (PMMA), polycarbonate (PC), polystyrene (PS) Or glass etc. That is, the material of the chip body 100 may be PMMA, PC, PS or glass.

The cover 180 may be a flat plate structure to facilitate the 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 upper surface of the chip body 100 . As shown in FIG. 1 , the cover 180 covers the entire upper surface of the chip body 100 ; the size of the bottom surface of the cover 180 may also be smaller than the size of the upper surface of the chip body 100 , and the cover 180 covers the part of the chip body 100 The upper surface is to cover the functional area of the chip body 100 to reduce the volume of the cover 180 . Of course, the cover 180 can also adopt other existing structures.

The cover 180 can be injection-molded, and the cover 180 and the chip body 100 are connected by bonding, such as thermal bonding, adhesive bonding, and ultrasonic bonding. The cover 180 may also be a film layer formed on the chip body 100 by a film sticking 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. The connection method between the cover 180 and the chip body 100 may also adopt other existing methods, which will not be repeated here.

The chip body 100 can also be injection molded. When the chip body 100 is injection-molded, a mold for the chip body 100 needs to be fabricated first, and the mold can be formed by electroforming, machining, or etching. The above-mentioned chip body 100 may also be manufactured by using other micro-manufacturing techniques such as laser etching molding, photolithography molding, and the like.

In the above embodiment, in each screening unit, the width L1 of the accommodating cavity 131 is larger than the diameter of the target cell, and the width L2 of the screening channel 132 is smaller than the diameter of the target cell. Since 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. , which is the diameter value of the smallest cell in the target cell.

In some possible examples, when the target cells are circulating tumor cells, the diameter of the target cells is about 10-20 μm, the width of the screening channel 132 is less than 10 μm, such as 8 μm, and the width of the accommodating cavity 131 is greater than 10 μm, such as 20 μm . 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 from the screening channel 132, thereby separating the target cells from the sample solution, and trapped in the accommodating cavity 131 . Preferably, each accommodating cavity 131 captures and traps a single target cell.

The screening array 130 may be a groove formed on the end face of the chip body 100 , for example, a groove formed on the upper surface of the chip body 100 . Correspondingly, the accommodating cavity 131 is a cavity formed on the upper surface of the chip body 100 . slot. 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, as shown in FIG. 3 , the length of L1 . The screening channel 132 is a guide groove formed on the upper surface of the chip body 10 , the opening of the guide groove faces the cover 180 , and the width of the screening channel 132 is the distance between two opposite side walls of the guide groove, such as Figure 3 shows the length of L2.

Taking a plane parallel to the upper surface of the chip body 100 as a cross-section, the cross-sectional shape of the accommodating cavity 131 is a rectangle, a trapezoid, a semicircle, a U-shape or a parabola. The width of the outlet end of the accommodating cavity 131 is smaller than or equal to the width of the inlet end of the accommodating cavity 131 , and the width of the inlet end of the accommodating cavity 131 is larger than the diameter of the target cells for capturing and retaining the target cells.

The widths of the plurality of accommodating cavities 131 in each screening array 130 may or may not be consistent. When the screening array 130 is provided with accommodating cavities 131 with various widths, the accommodating cavities 131 with different widths can capture target cells with different diameters, and try to avoid accumulation of multiple target cells in one accommodating cavity 131, so that one accommodating cavity 131 can accommodate The cavity 131 tries to trap a target cell so as to facilitate the identification of the target cell.

In some possible examples, the width of the accommodating cavity 132 located in the middle of the screening array 130 is greater than the width of the accommodating cavity 132 located at the two ends of the screening array 130, and the widths of the other accommodating cavities 132 may be the same. The middle value of the widths of the two accommodating cavities 132 .

In other possible examples, along the direction from the middle to the end of the screening array 130 , the width of the accommodating cavity 131 gradually decreases. There are five accommodating cavities 131 from the left end to the middle as shown by the dotted line in FIG. 4 . That is, the width of the accommodating cavity 131 gradually increases along the flow direction of the liquid inlet groove 120 . This arrangement prevents the target cells from accumulating in the accommodating cavity 131 at the end of the screening array 130 and improves the utilization rate of the accommodating cavity 131 in the middle.

Continuing to refer to FIG. 2 , in each screening unit, the 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, for example, the included angle is 45°. This arrangement can make the sample solution in the liquid inlet groove 120 smoothly flow into the accommodating cavity 131 in the screening unit, reduce the vortex and backflow in the sample solution, thereby reducing the impact on the target cells and the deformation and extrusion in the screening unit. channel 132. The target cells are trapped in the accommodating cavity 131, and the non-target cells with smaller diameters can flow out through the screening channel 132, thereby improving the capture rate of the target cells.

As the degree of the included angle between the direction of flow of the accommodating cavity 131 and the direction of flow of the liquid inlet groove 120 decreases, the length of the screening unit increases, that is, between the liquid inlet groove 120 and the liquid outlet groove 140 The flow channel through which the sample solution flows becomes longer. In order to prevent non-target cells with smaller diameters from accumulating in the longer screening channel 132 and affecting the flow rate of the sample solution, referring to FIG. 3 , a buffer tank 133 may be formed at the outlet end of the screening channel 132 , namely the screening channel 132 and the buffer tank 133 connected.

The buffer groove 133 and the screening channel 132 are projected on a plane perpendicular to the diversion direction of the screening channel 132 , and the projected area of the orthographic projection of the buffer groove 133 is larger than that of the screening channel 132 . In this way, 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 smoothness of the screening channel 132 can be maintained. In addition, the buffer tank 133 can also reduce that the sample solution flowing out of the upstream screening channel 132 passes through the liquid outlet groove 140 and then flows into the downstream screening channel 132, which will backflush with the sample solution flowing out of the screening channel 132, resulting in non-existent Target cells accumulate in this selection channel 132 .

In the present embodiment, a buffer groove 133 is formed at the outlet end of each screening channel 132 , and the buffer groove 133 may be approximately in the shape of a Mitsubishi column. As shown by the dotted line in FIG. 3 , the plane formed by the removal of the sharp corners of the left side wall of the screening channel 132 is a part of the bottom of the buffer groove 133 . The buffer groove 133 may also be a quadrangular prism or a semi-cylindrical shape, and the specific shape of the buffer groove 133 is not limited herein. One side of the quadrangular prismatic buffer groove 133 communicates with the outlet end of the screening channel 132 , and the arc surface of the semi-cylindrical buffer groove 133 communicates with the outlet end of the screening channel 132 .

Continuing to refer to FIG. 2 , among the plurality of screening units in each screening array, each screening unit includes a accommodating cavity 131 and a screening channel 132 , and the centerline of the accommodating cavity 131 coincides with the centerline of the screening channel 132 , which can improve the The uniformity of the flow of the sample solution in the screening unit. The centerlines of each screening unit on the same side are parallel to each other, so that the flow direction of each screening unit in the screening array 130 is consistent, so as to improve the uniformity of the flow of the sample solution in the screening array 130 .

Among the multiple screening units of each screening array, at least one screening unit may be provided with two or more screening channels. That is, at least one screening unit may include one accommodating cavity 131 and at least two screening channels 132 . In this way, the screening path of the sample solution is increased. Compared with the screening unit provided with only one screening channel 132, the screening unit provided with multiple screening channels 132 increases the shunt path and increases the shunt flow. The flow rate of the sample solution is increased, the screening time of the sample solution is shortened, and the screening efficiency is improved.

In some possible examples, each screening unit includes one accommodating cavity 131 and two screening channels 132 . 5 and 6 , the inlet end of each screening channel 132 is communicated with the outlet end of the accommodating cavity 131 , the outlet end of each screening channel 132 is communicated with the liquid outlet groove 140 respectively, and the diversion end of each screening channel 132 The included angle between the direction and the flow direction of the liquid outlet groove 140 may be an acute angle, and/or the included angle between the flow direction of each accommodating cavity 131 and the flow direction of the liquid inlet groove 120 may be is an acute angle. In this way, as shown by the arrow in FIG. 6 , when the sample solution in the screening channel 132 converges in the liquid outlet groove 140 , the reverse impact of the sample solution in the liquid outlet groove 140 on the sample solution flowing out of the screening channel 132 is avoided. As a result, the flow is blocked, and the target cells are even washed away from the accommodating cavity 131 , thereby improving the capture rate of the accommodating cavity 131 for the target cells.

In some other possible examples, in each screening array 130, a part of the screening units includes two screening channels 132, and another part of the screening units includes three screening channels 132, and the screening channels 132 are further increased for shunting, improving the the screening efficiency. Referring to FIGS. 7 and 8 , along the extending direction of the liquid inlet groove 120 , the screening units including three screening channels 132 and the screening units including two screening channels 132 are alternately arranged in sequence. Wherein, in the screening unit containing two screening channels 132, the included angle between the flow guiding direction of each screening channel 132 and the flow guiding direction of the accommodating cavity 131 is an acute angle, so as to facilitate flow diversion.

In order to further improve the capture rate of the accommodating cavity 131 , the liquid inlet groove 120 and the liquid outlet groove 140 may both be serpentine. The liquid inlet groove 120 includes at least two linear channels 121 parallel to each other, as shown in FIG. 9 . In the embodiment of the present disclosure, the liquid inlet groove 120 includes eight straight channels 121 . In every two adjacent straight channels 121 , the outlet of one straight channel 121 communicates with the inlet of the first arc-shaped channel 122 , and the outlet of the first arc-shaped channel 122 communicates with the inlet of the other straight channel 121 . That is, the straight channel 121 and the first arc-shaped channel 122 are connected in sequence, and in this way, the liquid inlet groove 120 and the liquid outlet groove 140 can be folded to reduce the length of the cell screening chip.

The extension direction of the straight channel 121 in the liquid inlet groove 120 may be consistent with the length direction of the cell screening chip. In the example shown in FIG. 9 , the flow direction of the straight channel 121 is parallel to the length direction of the cell screening chip. The extension direction of the straight channel 121 in the liquid inlet groove 120 may also be consistent with the width direction of the cell screening chip. The number and extension direction of the straight channels 121 in the liquid inlet groove 120 are arranged according to the usage requirements of the cell screening chip.

The first arc-shaped channel 122 in the liquid inlet groove 120 may be a semi-circular arc, connecting two adjacent straight channels 121 for the sample solution to flow through. Part of the screening array 130 may be provided between the liquid inlet groove 120 and the liquid outlet groove 140 , or all of the screening array 130 may be provided.

In a possible example, as shown in FIG. 10 , screening arrays 130 are provided on both sides of the straight channel 121 of the liquid inlet groove 120 , and arc guides are provided on both sides of the first arc-shaped channel of the liquid inlet groove 120 The flow plate 122 conducts the flow of the sample solution.

In another possible example, as shown in FIG. 11 , screening arrays 130 are provided on both sides of the straight channel 121 and the first arc-shaped channel 122 of the liquid inlet channel 120 . When the arc-shaped channel 122 turns, under the action of centrifugation, the target cells gather to the side of the first arc-shaped channel 122 away from the center of the arc, and this side captures more target cells.

In another possible example, as shown in FIG. 12 , screening arrays 130 are provided on both sides of the first arc-shaped channel 122 of the liquid inlet groove 120 , and linear guides are provided on both sides of the straight channel 121 of the liquid inlet groove 120 The flow plate uses centrifugation to capture the target cells. This setting can reduce the processing difficulty of the cell screening chip. Exemplarily, the distance between two adjacent accommodating cavities 131 located inside the first arc-shaped channel 122 is the first distance, and between two adjacent accommodating cavities 131 located outside the first arc-shaped channel 122 The distance is the second distance. The first distance is smaller than the second distance and the inlet ends of the accommodating chambers 131 located on both sides of the first arc-shaped channel 122 are staggered. Each accommodating cavity 131 may communicate with two screening channels 132 to improve screening efficiency.

One or more of the above structures can be arranged between the liquid inlet groove 120 and the liquid outlet groove 140 . For example, a screening array 130 is provided on both sides of part of the linear channel 121 of the liquid inlet groove 120 , connecting the above-mentioned linear channels 121 . Arc-shaped baffles 122 are arranged on both sides of the first arc-shaped channel, and screening arrays 130 are arranged on both sides of another part of the linear channel 121 of the liquid inlet groove 120, connecting the two sides of the first arc-shaped channel of this part of the linear channel 121 A screening array 130 is also provided. Alternatively, a screening array 130 is provided between the liquid inlet groove 120 and the liquid outlet groove 140 , that is, the screening arrays 130 are provided on both sides of all the straight channels 121 and all the first arcuate channels 122 of the liquid inlet groove 120 .

In other possible examples, the liquid inlet groove 120 and the liquid outlet groove 140 may both be wavy lines. Referring to FIG. 13 , the liquid inlet groove 120 includes at least two second arc-shaped channels 124 connected in sequence, and a screening array 130 is provided on the side of each second arc-shaped channel 124 . That is, no straight channel is provided in the liquid inlet groove 120 . In this way, 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 and reduce the accumulation of target cells.

The wavy line can be formed by connecting at least two semicircles in sequence. When the wavy line 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 can also be a sine curve or a cosine curve.

In some other possible examples, the liquid inlet groove 120 and the liquid outlet groove 140 may both be linear. Referring to FIGS. 14 and 15 , the direction from the middle of the liquid inlet groove 120 to the end of the liquid inlet groove 120 is shown in FIG. 14 and FIG. 15 . , the width of the liquid inlet groove 120 gradually increases. The left end of a section of liquid inlet groove 120 shown in FIG. 15 is close to the middle of the entire liquid inlet groove 120 , and the right end of a section of liquid inlet groove 120 shown in FIG. 15 is close to the end of the entire liquid inlet groove 120 . The width of the shown section of the liquid inlet groove 120 gradually increases from left to right.

That is, along the flow direction of the sample solution, the width of the entire liquid inlet groove 120 first decreases, then increases, and then decreases again. For example, the width of the liquid inlet groove 120 changes linearly from the end portion to the middle portion of the liquid inlet groove 120 . Taking a plane perpendicular to the center line of the liquid inlet groove 120 as a cross section, the cross-sectional area of the middle of the liquid inlet groove 120 is smaller than that of the end of the liquid inlet groove 120 . The flow rate in the middle of the liquid inlet groove 120 is greater than the flow rate at the end, so the side wall pressure in the middle of the liquid inlet groove 120 is greater than the side wall pressure at the end, so that the target cells in the sample solution enter the screening unit in the middle, and the pressure in the middle is improved. The capture rate of the filter unit. In this way, the accumulation of target cells in the screening unit at the end is reduced, the utilization rate of the screening unit in the middle is improved, and the target cells can be dispersed and trapped in multiple screening units in the screening array, so that most of the screening units can be captured. target cells for subsequent identification of target cells.

2, the chip body 100 shown in FIG. 2 is provided with a sample inlet 110, the sample inlet 110 communicates with the liquid inlet groove 120, and 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 before entering the liquid inlet groove 120 from the sample inlet 110 to separate the target cells. The sample solution contains various blood cells, such as white blood cells and red blood cells.

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 dual injection ports. Referring to FIG. 16 , the injection port 110 may include a first injection port 111 and a second injection port 112, wherein the first injection port 110 may be used for blood injection, or other liquid injections such as fixatives, staining solutions, 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.

The first injection port 111 and the second injection port 112 are respectively communicated with the liquid inlet groove 120 , and the blood sample entered through the first injection port 111 and the diluent entered through the second injection port 112 are in the liquid inlet groove 120 Mix while flowing. By controlling the flow rate of the blood sample and the diluent, the dilution ratio of the blood sample is controlled, and the required sample solution is formed in the liquid inlet groove 120 . This setting eliminates the need to dilute the blood sample in advance, shortens the detection time of target cells, and improves detection efficiency.

Between the sample inlet 110 and the liquid inlet groove 120, a liquid inlet connection pipe 160 may also be provided, and the liquid inlet connection pipe 160 is used for flow splitting, so that the sample inlet 110 can be connected to a plurality of liquid inlet grooves 120. On the one hand, The flow rate of the sample solution in the chip body 100 can be increased, thereby shortening the detection time. On the other hand, multiple screening arrays corresponding to the liquid inlet grooves 120 can capture target cells in parallel, thereby improving the detection efficiency.

Along the direction from the sample inlet 110 to the liquid inlet groove 120, the liquid inlet connection pipeline 160 includes at least two stages of liquid inlet split pipes arranged in sequence, and each stage of the liquid inlet split pipeline includes at least two liquid inlet split pipes 161 arranged in parallel. . Each liquid inlet split pipe 161 located in the upper stage is communicated with at least two liquid inlet split pipes 161 located in the next stage, and the first-level liquid inlet split pipe close to the injection port 110 is communicated with the injection port 110, and is adjacent to the injection port 110. The first-stage liquid inlet branch pipes of the liquid groove 120 are respectively communicated with the liquid inlet groove 120 .

The upper stage refers to the first stage in the adjacent two-stage liquid inlet split pipelines that is close to the injection port 110 , that is, the first stage located upstream along the flow direction of the sample solution. The next stage refers to the stage close to the liquid inlet groove 120 in the adjacent two-stage liquid inlet split pipes, that is, the stage located downstream along the flow direction of the sample solution.

In some possible examples, referring to FIG. 17 , the liquid inlet connection pipe 160 includes two-stage liquid inlet split pipes. For the convenience of description, the two-stage liquid inlet split pipes are respectively defined as the first stage liquid inlet split pipe 162 and the second stage liquid split pipe. Inlet shunt pipe 163. Wherein, the first-stage liquid inlet and branch pipes 162 are located on the left side as shown in FIG. 17 , and are the upper stage of the two-stage liquid inlet and branch pipes. The second-stage liquid inlet branch pipe 163 is located on the right side as shown in FIG. 17 , and is the next stage in the two-stage liquid inlet branch pipe.

The first-stage liquid inlet shunt pipeline 162 includes two liquid inlet shunt pipelines 161 arranged in parallel. The inlet ends of the two liquid inlet shunt pipelines 161 are communicated with each other, and both are communicated with the injection port 110. The two liquid inlet shunt pipelines The outlet port of 161 is not connected.

The second-stage liquid inlet shunt pipeline 163 includes four liquid inlet shunt pipelines 161 arranged in parallel, which can be divided into two groups. The inlet ends of the two liquid inlet shunt pipes 161 located above as shown in FIG. 19 are communicated with the outlet end of one of the two liquid inlet shunt pipes 161 of the first stage liquid inlet shunt pipe 162 . The inlet ends of the two liquid inlet split pipes 161 located below as shown in FIG. 19 are communicated with the outlet end of the other of the two liquid inlet split pipes 161 of the first stage liquid inlet split pipe 162 . The outlet ends of the four liquid inlet branch pipes 161 of the second stage are respectively communicated with one liquid inlet groove 120 .

Each liquid inlet branch pipe 161 located in the upper stage communicates with two liquid inlet branch pipes 161 located in the next stage. Taking the plane perpendicular to the centerline of the liquid inlet and branch pipes 161 as the cross section, the cross-sectional area of the liquid inlet branch pipes 161 located at the next stage is half of the cross-sectional area of the liquid inlet branch pipes 161 located at the previous stage. This arrangement can improve the uniformity of the flow of the sample solution in the liquid inlet grooves 120, so that the pressure in each liquid inlet groove 120 is balanced, and the pressure in a certain liquid inlet groove 120 is prevented from being too large, resulting in damage to the chip body 100. Work.

In other possible examples, referring to FIG. 18 , the liquid inlet connection pipe 160 includes a three-stage liquid inlet branch pipe. The first-stage liquid inlet splitting pipeline near the sample inlet 110 includes two liquid inlet splitting pipelines 161 arranged in parallel. The outlet ends of the two liquid inlet branch pipes 161 are not connected.

The first-stage liquid inlet splitting pipeline located in the middle includes four liquid inlet splitting pipelines 161 arranged in parallel, and the inlet ends of the two liquid inlet splitting pipelines 161 located at the top as shown in FIG. The outlet end of one liquid inlet split pipe 161 is connected, and the inlet ends of the two liquid inlet split pipes 161 located at the bottom shown in FIG. are connected, and the outlet ends of the four liquid inlet branch pipes 161 are not connected.

The first-stage liquid inlet branch pipes close to the liquid inlet groove 120 include eight liquid inlet branch pipes 161 arranged in parallel. The eight liquid inlet shunt pipes 161 are divided into four groups with two adjacent ones as a group, and the inlet ends of the two liquid inlet shunt pipes 161 in each group are communicated with each other and are respectively connected with the first-level liquid inlet shunt pipes in the middle. The outlet ends of each of the liquid inlet branch pipes 161 are communicated with each other. The outlet ends of the eight liquid inlet branch pipes 161 are respectively communicated with the liquid inlet grooves 120 . Taking the plane perpendicular to the center line of the liquid inlet shunt pipeline 161 as the cross section, in the adjacent two-stage liquid inlet shunt pipelines, the cross-sectional area of the liquid inlet shunt pipeline located at the next level is the section of the liquid inlet shunt pipeline located at the upper stage. half of the area.

A liquid outlet connection pipe 170 may also be provided between the liquid outlet groove 140 and the sample outlet 150 . The liquid outlet connecting pipe 170 is used for confluence, and the sample solutions of the plurality of liquid outlet grooves 140 are collected to the sample outlet 150 . Referring to FIGS. 17 and 18 , along the direction from the sample outlet 150 to the liquid outlet groove 140 , the liquid outlet connection pipe 170 includes at least two levels of liquid outlet branch pipes arranged in sequence. Each level of liquid outlet branch pipes includes at least two liquid outlet branch pipes 171 arranged in parallel, and each liquid outlet branch pipe 171 in the upper stage communicates with at least two liquid outlet branch pipes 171 in the next stage. The first-stage liquid outlet shunt pipeline close to the sample outlet 150 is communicated with the sample outlet 150 , and the first-stage liquid outlet shunt pipelines adjacent to the liquid outlet groove 140 are respectively communicated with the liquid outlet groove 140 . For the specific structure of the liquid outlet connecting pipe 170, reference may be made to FIG. 17 and FIG. 18 and the structure of the liquid inlet connecting pipe 160, which will not be repeated here.

It should be noted that, the chip body 100 may be provided with 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. The number of stages and the number of stages of the liquid outlet connection pipeline 170 may be the same or different. For example, the liquid inlet connection pipeline 160 includes two stages, and the liquid outlet connection pipeline 170 includes four stages. When the number of stages of the liquid inlet connection pipes 160 in the chip body 100 is the same as that of the liquid outlet connection pipes 170, the flow uniformity of the sample solution is better.

In the cell screening chip provided in the embodiment of the present disclosure, the chip body 100 includes a liquid inlet groove 120 , and along the flow direction of the liquid inlet groove 120 , the end of the liquid inlet groove 120 is closed, and the two sides of the liquid inlet groove 120 are closed. A liquid outlet groove 140 is provided, and a screening array 130 is arranged between the liquid inlet groove 120 and each liquid outlet groove 140 , and is communicated by the screening array 130 . The screening array 130 includes a plurality of screening units, each screening unit includes a accommodating cavity 131 and a screening channel 132 , and the liquid inlet groove 120 , the accommodating cavity 131 , the screening channel 132 and the liquid outlet groove 140 are connected in sequence. The sample solution containing the target cells flows from the liquid inlet groove 120 into the accommodating cavity 131 and the screening channel 132 in sequence, and flows out from the liquid outlet groove 140 . Because the width of the accommodating cavity 131 is greater than the diameter of the target cell, the width of the screening channel 132 is smaller than the diameter of the target cell, and the target cells cannot pass through the screening channel 132 and are trapped in the accommodating cavity 131 . The target cells flow out from the screening channel 132 to achieve the separation of target cells and non-target cells. Meanwhile, in the two screening arrays 130 , the inlet end of the accommodating cavity 131 in one of the screening arrays 130 and the inlet end of the accommodating cavity 131 in the other screening array 130 are arranged in a different position. Compared with the symmetrical distribution of the accommodating cavities on both sides of the liquid inlet groove, in the embodiment of the present disclosure, the accommodating cavities 131 on both sides of the liquid inlet groove 120 are staggeredly distributed, so that the accommodating cavities on both sides of the liquid inlet groove 120 are staggered. The cavities 131 are staggered, and the target cells only flow through one accommodating cavity 131 at a time, which is affected by the lateral flow rate in one direction, which reduces the probability that the target cells miss the accommodating cavity 131, improves the retention rate of the target cells, and further improves the capture rate of target cells.

Embodiment 2

Referring to FIG. 19 , an 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 sample injection pump 20 and a waste liquid collection device 70. The sample inlet of the cell screening chip 10 is connected to the sample injection pump 20, and the sample outlet is connected to the waste liquid collection device 70. The sampling 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. liquid.

The sampling pump 20 includes a sample solution pump 22 that pumps the sample solution into the cell screening chip 10 . In some possible examples, the sample solution pump 22 includes a blood sample pump and a diluent pump, and the blood sample and the diluent are mixed in the cell screening chip 10 through the structure of dual injection ports, thereby reducing the time required for detection. In other possible examples, the sample solution pump 22 pumps the diluted blood sample. Wherein, the diluent may be Phosphate Buffer Saline (PBS for short).

The sampling pump 20 may further include one or more of a surface treatment liquid pump 21 , a buffer liquid pump 23 , a fixative liquid pump 24 , and a staining liquid pump 25 . As shown in FIG. 19 , the sample injection pump 20 includes a treatment solution 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 .

When the sampling pump 20 includes a variety of pumps, a reversing valve 30 is disposed between the sampling pump 20 and the cell screening chip 10 . That is, the output end of the sampling 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, and the liquid in each pump is allowed to enter the cells in a certain order through the reversing valve 30. Screening chip 10.

It should be noted that the surface treatment liquid may be polyvinyl pyrrolidone (Polyvinyl Pyrrolidone, PVP for short), which is used to reduce the flow resistance of the sample solution. The buffer can be the same as the diluent, also PBS. The fixative solution can be a solution of 4% paraformaldehyde (Paraformaldehyde, PFA for short), which is used for stereotyping the target cells. Specifically, the fixative solution can reduce the elasticity of the cells, the cells after the action of the fixative solution are not easily deformed, and various structures in the cells are also fixed to realize the stereotype of the cells themselves. The staining solution can be a fluorescent stain for staining the target cells for easy identification, for example, when the target cells are circulating tumor cells, the fluorescent stain can include CD45, 4',6- _{difluorescein} with a fluorescein Amidino-2-phenylindole (4',6-diamidino-2-phenylindole, referred to as DAPI) and epithelial cell adhesion molecule (Epithelial Cell Adhesion Molecule, referred to as EpCAM) with another fluorescein, among which, CD ₄₅ is used to label leukocytes, EpCAM is used to label circulating tumor cells, and DAPI is used to label cell nuclei. Target cells can be further identified by staining different cells.

Continuing to refer to FIG. 19 , the cell screening system further includes a light source 40 , an image acquisition device 50 and a data processing device 60 . Wherein, the light source 40 is used to illuminate the cell screening chip 10 when the image acquisition device 50 is working, and the light source 40 may 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 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 respectively. 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 selection chip 10 .

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

Continuing to refer to FIG. 19 , the cell screening system may further include a stage 80 for placing the cell screening chip 10 . The stage 80 can be a conveyor belt, so that the cell screening chip 10 moves relative to the image acquisition device 50 at a certain speed, so as to ensure that the image acquisition device 50 can acquire an image of the entire cell screening chip 10 .

In the embodiment of the present disclosure, the cell screening system includes a sample injection pump 20 , a waste liquid collection device 70 and the above-mentioned cell screening chip 10 . The port is connected to the waste liquid collection device 70. The cell screening system includes the above cell screening chip, so it has the advantage of high capture rate of target cells. The specific effect can be referred to above, and will not be repeated here.

Embodiment 3

Referring to FIG. 20 , an embodiment of the present disclosure provides a cell screening method, which is applicable to the above-mentioned cell screening system for separating and identifying target cells, and the cell screening method includes:

S101. The cell screening chip 10 is sequentially injected with a surface treatment solution and a buffer solution for pretreatment.

The surface treatment liquid is pumped into the cell screening chip 10 through the sampling 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 solution is pumped into the cell screening chip 10 through the sampling pump 20, the surface treatment liquid is rinsed, and the cell screening chip 10 is filled with air bubbles.

S102, inject the sample solution containing the target cells into the cell screening chip 10, and the cell screening chip 10 captures the target cells.

In the embodiment of the present disclosure, the diluted blood sample can be pumped into the cell screening chip 10 through the sample injection pump 20 , that is, the mixed sample solution is pumped into the cell selection chip 10 , or the blood samples can be pumped separately through the sample injection pump 20 . The sample solution is formed after mixing with the diluent into the cell screening chip 10 .

The cell screening chip 10 is used to separate the target cells from the sample solution, the non-target cells with a smaller size flow out of the cell screening chip 10, and the target cells with a larger size are captured and retained by the cell screening chip 10, thereby separating the target cells from the non-target cells. Cell isolation. The cell screening chip 10 is the cell screening chip 10 described above, and details are not repeated here. The cell screening chip 10 in the embodiment of the present disclosure has a high capture rate for target cells.

It should be noted that the material of the cell screening chip 10 may be a transparent material, such as glass, so as to facilitate subsequent identification of the target cells in the cell screening chip 10 .

S103, injecting buffer solution, fixative solution, buffer solution, staining solution and buffer solution into the cell screening chip 10 in sequence, to finalize and stain the target cells.

In the embodiment of the present disclosure, the buffer solution is pumped into the cell selection chip 10 through the sampling pump 20 to clean the cell selection chip 10; the fixative solution is pumped into the cell selection chip 10 through the sampling pump 20 to finalize the target cells , the fixative can be PFA; the buffer solution is pumped into the cell screening chip 10 by the sampling pump 20, the cell screening chip 10 is cleaned again, and the fixative is washed away; the dyeing solution is pumped by the sampling pump 20 to the cell screening In the chip 10, the target cells are marked to further distinguish the cell types in the cell screening chip 10, and the staining solution may be a fluorescent dye.

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 7-20μm. When the target cells are circulating tumor cells, the cell screening chip 10 will capture circulating tumor cells and some leukocytes. In order to distinguish the above two kinds of cells, the two kinds of cells can be marked with different fluorescent colors by the staining solution, so as to facilitate the identification of circulating tumor cells.

After staining, the sample injection pump 20 pumps the buffer into the cell screening chip 10 , cleans the cell screening chip 10 again, and rinses the staining solution to facilitate the subsequent collection of fluorescence images of the cell screening chip 10 .

S104 , the image acquisition device 50 acquires an 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.

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 fill light for the image acquisition device 50 , so that the image acquisition device 50 Clearer images can be captured.

The image acquisition device 50 is signal-connected to 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 sequentially injected with a surface treatment solution and a buffer for pretreatment, and then a sample solution is injected, and the target cells in the sample solution are captured by the cell screening chip 10, because the cell screening method is the above-mentioned cell screening method. The method corresponding to the screening system can improve the capture rate of the target cells, which will not be repeated here. At the same time, the fixative solution is used to finalize the target cells, and the staining solution is used to stain and identify the target cells to distinguish them from non-target cells, which is convenient for the identification of the target cells.

The embodiments or implementations in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments, and the same and similar parts between the various embodiments may be referred to each other.

It should be understood by those skilled in the art that in the disclosure of the present disclosure, the terms "portrait", "horizontal", "upper", "lower", "front", "rear", "left", "right", " The orientation or positional relationship indicated by vertical, horizontal, top, bottom, inner, outer, etc. is based on the orientation or positional relationship shown in the drawings, which are only for convenience in describing the present disclosure and to simplify the description, rather than to indicate or imply that the system or element referred to must have a particular orientation, be constructed and operate in a particular orientation, and thus the above terms should not be construed as limitations of the present disclosure.

In the description of this specification, references to the terms "one embodiment," "some embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples" and the like are meant to incorporate embodiments A particular feature, structure, material, or characteristic described or exemplified 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 to illustrate the technical solutions of the present disclosure, but not to limit them; although the present disclosure has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: The technical solutions described in the foregoing embodiments can still be modified, or some or all of the technical features thereof can be equivalently replaced; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the technical solutions of the embodiments of the present disclosure. scope.

Claims

A cell screening chip, characterized in that it includes a chip body, the chip body is provided with a liquid inlet groove, and along the flow direction of the liquid inlet groove, the end of the liquid inlet groove is closed, and the liquid inlet groove is closed. Two sides of the liquid groove are respectively provided with a liquid outlet groove, and a screening array is formed between the liquid inlet groove and each of the liquid outlet grooves;

Each of the screening arrays includes a plurality of screening units arranged along the diversion direction of the liquid inlet groove, each screening unit includes a accommodating cavity and a screening channel, and the inlet end of the accommodating cavity is connected to the liquid inlet. The groove is communicated, 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; and the width of the accommodating cavity is larger than that of the target cell. diameter, the width of the screening channel is smaller than the diameter of the target cell;

In the two screening arrays, the inlet end of the accommodating cavity in one of the screening arrays and the inlet end of the accommodating cavity in the other screening array are arranged in a staggered position.
The cell screening chip according to claim 1, wherein among the plurality of screening units included in each of the screening arrays, at least one of the screening units includes one of the accommodating chambers and at least two of the screening units. filter channel.
The cell screening chip according to claim 2, wherein among the plurality of screening units included in each of the screening arrays, each of the screening units includes one accommodating cavity and two of the screening channels .
The cell screening chip according to claim 3, wherein the included angle between the diversion direction of each of the screening channels and the diversion direction of the liquid outlet groove is an acute angle;

And/or, the included angle between the flow direction of each accommodating cavity and the flow direction of the flow direction in the liquid inlet channel is an acute angle.
The cell screening chip according to claim 3, wherein, taking a plane parallel to the upper surface of the chip body as a cross-section, the cross-sectional shape of the accommodating cavity is a rectangle, a trapezoid, a semicircle, a U-shape or Parabolic, and the width of the outlet end of the accommodating cavity is smaller than or equal to the width of the inlet end of the accommodating cavity, and greater than the diameter of the target cell.
The cell screening chip according to claim 5, wherein a buffer groove is formed at the outlet end of the screening channel, and the diversion direction of the buffer groove coincides with the diversion direction of the screening channel;

The orthographic projection of the buffer groove on a plane perpendicular to the diversion direction of the screening channel is greater than the orthographic projection of the screening channel on the plane.
The cell screening chip according to claim 1, wherein the width of the accommodating cavity located in the middle of the screening array is greater than the width of the accommodating cavity located at the end of the screening array.
The cell screening chip according to claim 7, wherein the width of the accommodating cavity gradually decreases along the direction from the middle to the end of the screening array.
The cell screening chip according to 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 connected to The liquid outlet grooves communicate with each other.
The cell screening chip according to claim 9, wherein the injection port comprises a first injection port and a second injection port respectively communicated with the liquid inlet groove.
The cell screening chip according to claim 9, wherein the chip body is further provided with a liquid inlet connection pipeline located between the liquid inlet groove and the sample inlet;

Along the direction from the sample inlet to the liquid inlet groove, the liquid inlet connection pipeline includes at least two stages of liquid inlet split pipelines arranged in sequence, and each stage of the liquid inlet split pipeline includes at least two parallel distribution pipelines. a liquid shunt pipeline, and each of the liquid inlet shunt pipelines located in the upper stage is communicated with at least two of the liquid inlet shunt pipelines located in the next stage;

And in the liquid inlet and split pipelines at all levels, the first level of the liquid inlet split pipeline closest to the injection port is connected with the injection port, and the first level of the liquid inlet split pipe closest to the liquid inlet groove is connected. The pipes are respectively communicated with the liquid inlet grooves.
The cell screening chip according to claim 9 or 11, wherein the chip body is further provided with a liquid outlet connecting pipe between the liquid outlet groove and the sample outlet;

Along the direction from the sample outlet to the liquid outlet groove, the liquid outlet connection pipeline includes at least two stages of liquid outlet split pipes arranged in sequence, and each level of the liquid outlet split pipeline includes at least two parallel outlet pipes. liquid shunt pipes; and each of the liquid outlet shunt pipes located in the upper stage communicates with at least two of the liquid outlet shunt pipes located in the next stage;

And in the liquid outlet split pipes of all levels, the liquid outlet split pipes of the first stage closest to the sample outlet are communicated with the sample outlet, and the liquid outlet split pipes of the first stage closest to the liquid outlet groove are connected. The pipes are respectively communicated with the liquid outlet grooves.
The cell screening chip according to any one of claims 1-11, wherein the liquid inlet groove and the liquid outlet groove are both serpentine, and the liquid inlet groove comprises at least two parallel to each other. two straight passages, in every two adjacent straight passages, the outlet of one of the straight passages is communicated with the inlet of the other straight passage through the first arc-shaped passage;

The screening array is provided on the side of the straight channel and/or the first arc-shaped channel.
The cell screening chip according to any one of claims 1-11, wherein the liquid inlet groove and the liquid outlet groove are both wavy lines, and the liquid inlet groove comprises at least Two second arc-shaped channels, each of which is provided with the screening array on the side of the second arc-shaped channel.
The cell screening chip according to claim 14, wherein the wavy line shape is formed by connecting semicircles in sequence, or the wavy line shape is one of a sine curve and a cosine curve.
The cell screening chip according to any one of claims 1 to 11, wherein the liquid inlet groove is linear, and the liquid inlet groove extends from the middle of the liquid inlet groove to the end of the liquid inlet groove. direction, the width of the liquid inlet groove gradually increases.
A cell screening system, characterized in that it comprises a sample injection pump, a waste liquid collection device and the cell screening chip according to any one of claims 1-16, wherein the sample inlet of the cell screening chip is connected to the sample injection port. The pump is connected, and the sample outlet of the cell screening chip is connected to the waste liquid collection device.
The cell screening system according to claim 17, wherein the sampling pump comprises a sample solution pump, and one or more of a surface treatment solution pump, a buffer solution pump, a fixative solution pump, and a staining solution pump;

The output end of the injection pump is connected to one end of the reversing valve, and the other end of the reversing valve is connected to the injection port.
The cell screening system according to claim 17, wherein the cell screening system further comprises a light source, an image acquisition device and a data processing device;

The light source is used for illuminating the cell screening chip when the image acquisition device is working;

The image acquisition device is signal-connected to the data processing device, and the image acquisition device acquires the image of the cell screening chip and transmits it to the data processing device;

The data processing device identifies the target cell according to the image.
A cell screening method, wherein the cell screening system according to any one of claims 17-19 is adopted, and the cell screening method comprises:

The cell screening chip was sequentially injected with surface treatment solution and buffer for pretreatment;

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

injecting buffer, fixative, buffer, staining solution and buffer into the cell screening chip in sequence, and finalizing and staining the target cells;

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