Fluid capture chip, single cell capture and transfer system, and single cell analysis method

Abstract

The present invention discloses a fluid capture chip, a single cell capture and transfer system and a single cell analysis method. The fluid capture chip comprises a horizontally arranged and flat-plate-shaped fluid capture chip body, the fluid capture chip body having a cell suspension inlet, and the lower end of the fluid capture chip body having a plurality of spaced-apart single cell capture structures, and the plurality of single cell capture structures are all connected to the cell suspension inlet, the cell suspension inlet is used for adding a single cell suspension containing a plurality of single cells, and the single cell capture structure is used for capturing any single cell in the single cell suspension, and the structure is simple, so that a single cell suspension can be added through the cell suspension inlet, and the single cell suspension is captured by the single cell capture structure when flowing through the single cell capture structure, and the plurality of single cell capture structures capture a plurality of independent single cells, thereby realizing efficient capture of a small number of single cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of single cell analysis, and in particular relates to a fluid capture chip, a single cell capture and transfer system, and a single cell analysis method. Background Art

[0002] Microfluidics has unique advantages in analyzing single cells. Its micron-scale pipelines and nanoliter-scale systems are very close to the scale of cells and the environment in which cells live in the body. In addition, its high-throughput and parallel array analysis further improves its credibility and efficiency. However, single-cell analysis technology has not been widely used due to its high technical threshold and complex operation.

[0003] At present, the research on individual cells, especially single-cell analysis technology, mainly uses optical tweezers technology, microdroplet technology, flow cytometry, dielectrophoresis technology and photoinduced dielectrophoresis technology, hydrodynamic capture technology, micropit (well) technology and single-cell printing technology. However, these methods have the following problems to a greater or lesser extent: 1) High randomness: The use of microdroplet technology can well encapsulate single cells and interrupt the mutual communication between cells, but this method is difficult to break through the bottleneck of Poisson distribution, and a large number of empty droplets will be generated, so the randomness is high and it is difficult to ensure that most units capture single cells; micropit technology uses the random sedimentation of cells into the capture site, and also faces the problem of high randomness; 2) Complex operation and equipment: The use of optical tweezers, dielectrophoresis and photoinduced dielectrophoresis for high-precision single-cell capture not only faces the problems of complex equipment, difficult processing design, and high cost, but also needs to consider the problems of complex operation and low throughput; high-throughput analysis of single cells using flow cytometry is expensive and requires regular maintenance, which is difficult for most laboratories and clinical testing institutions to afford; 3) Poor addressability and insufficient analysis depth: Using microdroplet technology and flow cytometry, single cells are encapsulated in microdroplets for high-throughput analysis. After the microdroplets pass through the detection site, it is difficult to analyze 4) Insufficient culture space: Using droplet technology and hydrodynamic capture, single cells are confined to a very small space after capture, making it difficult to obtain sufficient oxygen and nutrients. Metabolic waste is easy to accumulate, which inevitably affects the normal growth of cells. 5) Cross-contamination is easy: With traditional hydrodynamic capture technology and micropit (well) technology, as well as single-cell printing technology, cells are often exposed to the same solute, and there is the possibility of paracrine cell-to-cell communication between cells, which is not a single-cell analysis in the full sense. 6) Multi-cell events are prone to occur: In recent years, a number of dual-microwell structure microfluidic systems have been developed and launched, which even combine dielectrophoresis technology for single-cell capture and analysis. However, this type of method still inevitably causes multi-cell events due to differences in individual cell volume and fluid movement.

[0004] In summary, single-cell analysis has a wide range of applications in life sciences and medicine, and is of great significance and value. However, there are still many difficulties and challenges in the development of this technology; at the same time, the existing single-cell analysis technology and equipment also have many shortcomings. At present, the market is in urgent need of a microfluidic system that can perform high-throughput, addressable, microenvironment-controllable single-cell arrayed, parallelized deep analysis, and can perform efficient single-cell analysis on extremely small clinical samples. Summary of the invention

[0005] In order to solve the above technical problems, one of the objectives of the present invention is to provide a fluid capture chip that can be used to capture a single cell array from a suspension containing a small amount of single cells.

[0006] In order to achieve the above-mentioned purpose, the technical solution of the present invention is as follows: a fluid capture chip, comprising a horizontally arranged and flat-plate-shaped fluid capture chip body, the fluid capture chip body having a cell suspension inlet for adding a single-cell suspension containing multiple single cells, and the lower end of the fluid capture chip body having a plurality of spaced-apart single-cell capture structures, and the plurality of single-cell capture structures are all connected to the cell suspension inlet, and the single-cell capture structure is used to capture any single cell in the single-cell suspension.

[0007] The beneficial effects of the above technical solution are: its structure is simple, so that a single-cell suspension can be added through the cell suspension inlet, and the single-cell suspension is captured by the single-cell capture structure when flowing through the single-cell capture structure, and multiple single-cell capture structures capture multiple independent single cells, thereby achieving the separation of multiple single cells from the single-cell suspension.

[0008] The fluid capture chip body in the above technical solution also includes a culture medium inlet and a sample outlet horizontally spaced apart, and the culture medium inlet, cell suspension inlet and sample outlet all penetrate the fluid capture chip body from top to bottom, and the cell suspension inlet is close to the culture medium inlet. A microfluidic channel is etched at the lower end of the fluid capture chip body, and the two ends of the microfluidic channel are respectively connected to the culture medium inlet and the sample outlet, and the cell suspension inlet is connected to an end of the microfluidic channel close to the culture medium inlet. Multiple single-cell capture structures are etched at the lower end of the fluid capture chip body, and the single-cell capture structure has a liquid inlet and a liquid outlet. Multiple single-cell capture structures are connected in series on the microfluidic channel and are all located between the cell suspension inlet and the sample outlet.

[0009] The beneficial effect of the above technical solution is that its structure is simple, so that the microfluidic cell suspension is mixed with the culture medium, and when flowing through the single-cell capture structure, the single cell is captured by the single-cell capture structure. At this time, the single cell is in the environment of the culture medium and can maintain its activity throughout the process while carrying out metabolism.

[0010] In the above technical solution, there are multiple microfluidic channels, and each of the microfluidic channels located between the cell suspension inlet and the sample outlet has multiple single-cell capture structures connected in series.

[0011] The beneficial effect of the above technical solution is that its structure is simple, so that the number of single cell capture structures is greater and more single cells can be separated.

[0012] In the above technical solution, the plurality of single cell capture structures are distributed in an array at the lower end of the fluid capture chip body.

[0013] The beneficial effect of the above technical solution is that it is more neat and beautiful.

[0014] The single-cell capture structure in the above technical solution is a groove horizontally etched at the lower end of the fluid capture chip body and having a "U"-shaped horizontal cross-section. The two ends of the single-cell capture structure are its liquid inlet and liquid outlet respectively, and the middle bend is a bend. A trumpet-shaped notch is provided between the liquid inlet and the bend, and the thin end of the notch is connected with the area between the bend and the liquid outlet, and the notch is used to capture single cells.

[0015] The beneficial effects of the above technical solution are: its structure is simple and its capture performance is good. When capturing single cells, a carrier plate needs to be placed at the lower end of the fluid capture chip body to seal the lower end of the fluid capture chip body and allow the microfluidic channel and the single cell capture structure to form a fluid passage under the enclosure of the carrier plate.

[0016] The second object of the present invention is to provide a single cell capture and transfer system which has a simple structure and good separation effect and can separate single cells.

[0017] In order to achieve the above-mentioned purpose, the technical scheme of the present invention is as follows: a single-cell capture and transfer system, comprising a through-glass chip, a microgroove array chip and the fluid capture chip as described above, wherein the fluid capture chip, the through-glass chip and the microgroove array chip are horizontally arranged in sequence from top to bottom and overlap with each other, and the through-glass chip has a plurality of microholes running through it from top to bottom; the microgroove array chip is concavely provided with a plurality of microgrooves, and the plurality of micropores, the plurality of microgrooves and the plurality of single-cell capture structures correspond to each other one by one, and each of the micropores is aligned with the corresponding microgroove, and the fluid capture chip can be horizontally moved at the upper end of the through-glass chip under the action of external force until the single-cell capture structure thereon is staggered with the corresponding micropore to capture a single cell, or moved to align with each other to discharge the captured single cell through the corresponding micropore into the corresponding microgroove.

[0018] The beneficial effect of the above technical solution is that: the fluid capture chip can be used in a state of being misaligned with the glass through-hole chip, and each single-cell capture structure of the fluid capture chip can capture a single cell from the fluid respectively. Then, when the fluid capture chip and the glass through-hole chip are aligned, the single cell captured by the single-cell capture structure is transferred downward through the corresponding micropore to the corresponding microgroove of the microgroove array chip, and finally there is only one cell in each microgroove of the microgroove array chip, thereby achieving a single-cell microarray.

[0019] The upper surface of the microgroove array chip in the above technical solution is provided with a hydrophobic layer.

[0020] The beneficial effect of the above technical solution is that: in this way, the microgroove array chip only has the fluid containing the single cell in the microgroove, and the upper surface of the microgroove array chip remains clean.

[0021] The hydrophobic layer in the above technical solution is a super hydrophobic film.

[0022] The beneficial effect of the above technical solution is that the upper surface of the microgroove array chip is made cleaner without any residual liquid.

[0023] A third object of the present invention is to provide a single cell analysis method using the single cell capture and transfer system as described above, characterized in that it comprises the following steps:

[0024] Step 1: Take a fluid capture chip and a through-glass chip, and stack the fluid capture chip on the through-glass chip in an offset manner;

[0025] Step 2: adding culture medium to the culture medium inlet of the fluid capture chip, and then taking a cell suspension using a capillary and inserting it into the cell suspension inlet, the cell suspension in the capillary flows along with the culture medium in the microfluidic channel toward the sample outlet, and the single cell in the culture medium is captured by the single cell capture structure;

[0026] Step 3: When the culture medium flows to the sample outlet, take the microgroove array chip and stack it on the lower end of the through-glass chip, and align each of the microholes of the through-glass chip with the corresponding microgrooves on the microgroove array chip;

[0027] Step 4: horizontally sliding the fluid capture chip until its single cell capture structures are aligned with the corresponding micropores, so as to transfer the single cells captured by the single cell capture structures to the corresponding microgrooves through the corresponding micropores;

[0028] Step 5: After the single cell transfer in step 4 is completed, the microgroove array chip is placed in a culture dish for single cell culture.

[0029] The beneficial effect of the above technical solution is that the method is simple and can ensure that each single cell capture structure can capture at most one single cell, so that at most one cell will appear in each microgroove, and it can flexibly realize the capture of single cells.

[0030] A fourth object of the present invention is to provide a method for using the single cell capture and transfer system or the single cell analysis method as described above in drug screening. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a top view of the fluid capture chip described in Example 1 of the present invention;

[0032] Figure 2 This is an enlarged view of the single cell capture structure described in Example 1 of the present invention;

[0033] Figure 3 This is a vertical cross-sectional view of the fluid capture chip described in Example 1 of the present invention;

[0034] Figure 4 A state diagram of the alignment of the fluid capture chip and the glass through-hole chip in the single-cell microfluidic system described in Example 2 of the present invention;

[0035] Figure 5 This is a state diagram of the fluid capture chip and the glass through-hole chip in the single-cell microfluidic system described in Example 2 of the present invention when they are misaligned;

[0036] Figure 6 It is a front view of the through glass via chip described in Example 2 of the present invention;

[0037] Figure 7 is a vertical view of the microgroove array chip described in Example 2 of the present invention;

[0038] Figure 8 This is an enlarged view of the cell suspension injection port described in Example 2 of the present invention.

[0039] In the figure: 1 fluid capture chip body, 11 cell suspension inlet, 12 single cell capture structure, 13 culture medium inlet, 14 sample outlet, 15 microfluidic channel, 121 liquid inlet, 122 liquid outlet, 123 bend, 124 notch, 2 glass through-hole chip, 21 micropore, 3 microgroove array chip, 31 microgroove. DETAILED DESCRIPTION

[0040] The principles and features of the present invention are described below in conjunction with the accompanying drawings. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0041] Example 1

[0042] like Figure 1-Figure 3As shown, this embodiment provides a fluid capture chip, including a horizontally arranged and flat-plate-shaped fluid capture chip body 1, the fluid capture chip body 1 having a cell suspension injection port 11, and the lower end of the fluid capture chip body 1 having a plurality of spaced single cell capture structures 12, and the plurality of single cell capture structures 12 are all connected to the cell suspension injection port 11, the cell suspension injection port 11 is used to add a single cell suspension containing a plurality of single cells, and the single cell capture structure 12 is used to capture any single cell in the single cell suspension, and its structure is simple, so that the single cell suspension can be added through the cell suspension injection port, and the single cell suspension is captured by the single cell capture structure when flowing through the single cell capture structure, and the plurality of single cell capture structures capture a plurality of independent single cells, thereby realizing the separation of a plurality of single cells from the single cell suspension. Among them, the aperture of the cell suspension injection port is small enough to meet the quantitative capillary insertion.

[0043] The fluid capture chip body 1 in the above technical solution also includes a culture medium inlet 13 and a sample outlet 14 that are horizontally spaced, and the culture medium inlet 13, the cell suspension inlet 11 and the sample outlet 14 all penetrate the fluid capture chip body 1 from top to bottom, and the cell suspension inlet 11 is close to the culture medium inlet 13. A microfluidic channel 15 is etched at the lower end of the fluid capture chip body 1, and the two ends of the microfluidic channel 15 are respectively connected to the culture medium inlet 13 and the sample outlet 14, and the cell suspension inlet 11 and the microfluidic channel 15 are close to the culture medium inlet 13. One end of the microfluidic chip body 1 is connected, and the multiple single-cell capture structures 12 are etched on the lower end of the fluid capture chip body 1, and the single-cell capture structure 12 has a liquid inlet 121 and a liquid outlet 122. The multiple single-cell capture structures 12 are connected in series on the microfluidic channel 15 and are located between the cell suspension inlet 11 and the sample outlet 14. The structure is simple, so that the microfluidic cell suspension is mixed with the culture medium, and when flowing through the single-cell capture structure, the single cell is captured by the single-cell capture structure. At this time, the single cell is in the environment of the culture medium, and it can remain active during the whole process and metabolize at the same time.

[0044] In the above technical solution, there are multiple microfluidic channels 15, and each of the microfluidic channels 15 located between the cell suspension inlet 11 and the sample outlet 14 is respectively connected in series with multiple single-cell capture structures 12. The structure is simple, so that the number of single-cell capture structures is greater and more single cells can be separated.

[0045] In the above technical solution, the plurality of single cell capture structures 12 are distributed in an array at the lower end of the fluid capture chip body 1, which is more neatly processed and beautiful.

[0046] The single-cell capture structure 12 in the above technical solution is a groove horizontally etched at the lower end of the fluid capture chip body 1 and having a "U"-shaped horizontal cross-section. The two ends of the single-cell capture structure are the liquid inlet 121 and the liquid outlet 122, respectively. The middle bend is a bend 123, and a trumpet-shaped notch 124 is provided between the liquid inlet 121 and the bend 123. The thin end of the notch (i.e., the end with a smaller opening) is connected to the area between the bend 123 and the liquid outlet 122, and the notch 124 is used to capture single cells. It has a simple structure and good capture performance. When capturing single cells, a carrier plate needs to be placed at the lower end of the fluid capture chip body to seal the lower end of the fluid capture chip body and allow the microfluidic channel and the single-cell capture structure to form a fluid passage under the enclosure of the carrier plate.

[0047] Example 2

[0048] like Figure 4-Figure 7 As shown, this embodiment provides a single-cell microfluidic system, including a fluid capture chip, a glass through-hole chip 2 and a microgroove array chip 3 which are horizontally arranged from top to bottom and overlapped with each other, wherein the fluid capture chip is as shown in Example 1, the glass through-hole chip 2 has a plurality of microholes 21 running through it from top to bottom; the microgroove array chip 3 is concavely provided with a plurality of microgrooves 31, and the plurality of microholes 21, the plurality of microgrooves 31 and the plurality of single-cell capture structures 12 correspond to each other one by one, each of the microholes 21 and the corresponding microgroove 31 are aligned with each other, and the fluid capture chip can be horizontally moved to the upper end of the glass through-hole chip 2 to the single cell thereon under the action of an external force. The cell capture structure 12 is staggered with the corresponding micropore 21 to capture a single cell, or moved to align with each other to discharge the captured single cell through the corresponding micropore 21 into the corresponding microgroove 31. In this way, the fluid capture chip can be used in a state where it is staggered with the glass through-hole chip, and each single cell capture structure of the fluid capture chip can capture a single cell from the fluid respectively, and then the fluid capture chip and the glass through-hole chip are aligned, so that the single cell captured by the single cell capture structure is transferred downward through the corresponding micropore to the corresponding microgroove of the microgroove array chip, and finally there is only one cell in each microgroove of the microgroove array chip, thereby obtaining multiple single cells. Wherein, the glass through-hole chip constitutes the carrier described in Example 1.

[0049] The upper surface of the microgroove array chip 3 in the above technical solution is provided with a hydrophobic layer, so that only the microgrooves of the microgroove array chip contain fluid containing single cells, and the upper surface of the microgroove array chip remains clean.

[0050] Preferably, the hydrophobic layer in the above technical solution is a super-hydrophobic film, so that the upper surface of the microgroove array chip is more clean and no liquid remains.

[0051] Wherein, preferably, the fluid capture chip, the glass through hole chip 2 and the microgroove array chip 3 are of the same size and are all rectangular plates, and further preferably, when the sides of the three are aligned, each notch is aligned with the corresponding micropore and microgroove. Wherein, the principle of the single cell capture structure capturing a single cell is: when the fluid capture chip and the glass through hole chip are misaligned, the lower ends of the microfluidic channel and the single cell capture structure are blocked by the glass through hole chip, and the microfluidic channel and the single cell capture structure are combined to form a fluid channel, and the fluid containing the single cell flows to the notch, and the thin end of the notch is in a negative pressure state due to the action of the fluid at the liquid outlet, and the single cell is attracted to the thin end of the notch, but the size of the single cell is larger than the size of the thin end of the notch, causing the single cell to be intercepted at the thin end of the notch, so as to achieve the capture of the single cell, and when the fluid capture chip is aligned with the glass through hole chip, each notch is connected to the corresponding microgroove through the corresponding micropore, and the fluid at the notch drives the intercepted single cell to flow downward into the microgroove along with the corresponding micropore, thereby realizing the transfer of the intercepted single cell into the microgroove.

[0052] After the single cell captured by the single cell capture structure is transferred to the microgroove, a trace amount of culture medium is transferred along with the single cell. At this time, the microgroove where the single cell is transferred also contains culture medium, so that the single cell has a good growth and development environment in the microgroove.

[0053] Among them, preferably, Figure 8 As shown, the cell suspension injection port is trumpet-shaped, and its larger opening end faces upward, so as to preferably meet the requirements of quantitative capillary insertion.

[0054] Example 3

[0055] This embodiment provides a single cell analysis method using the single cell capture and transfer system described in Example 2, which comprises the following steps:

[0056] Step 1: Take a fluid capture chip and a through-glass chip 2, and stack the fluid capture chip on the through-glass chip 2 in an offset manner;

[0057] Step 2: adding culture medium to the culture medium inlet 13 of the fluid capture chip, and then taking a cell suspension using a capillary and inserting it into the cell suspension inlet 11; the cell suspension in the capillary flows along with the culture medium in the microfluidic channel 15 toward the sample outlet 14, and the single cell in the culture medium is captured by the single cell capture structure 12;

[0058] Step 3: When the culture medium flows to the sample outlet 14, take the microgroove array chip 3 and stack it on the lower end of the through-glass chip 2, and align each of the microholes 21 of the through-glass chip 2 with the corresponding microgrooves 31 on the microgroove array chip 3;

[0059] Step 4: horizontally slide the fluid capture chip until its single cell capture structure 12 is aligned with the corresponding microhole 21, so as to transfer the single cell captured by the single cell capture structure 12 to the corresponding microgroove 31 through the corresponding microhole 21;

[0060] Step 5: After the single cell transfer in step 4 is completed, the microgroove array chip 3 is placed in a culture dish for single cell culture. This method is simple and can ensure that each single cell capture structure can capture only one single cell at most, so that only one cell will appear in each microgroove at most, which can flexibly realize the capture of single cells.

[0061] Wherein, in the step 2, before adding the culture medium, in order to avoid bubbles in the microfluidic channel, acetonitrile can be added from the culture medium inlet first, and after the acetonitrile fills the microfluidic channel, PBS buffer is introduced from the culture medium inlet to rinse the acetonitrile in the microfluidic channel, and then the culture medium is added. Preferably, the upper end of the fluid capture chip is located at the sample outlet and can be connected to a drainage tube, so that the drainage tube can be used to divert the material discharged from the sample outlet to avoid the material contaminating the upper end of the fluid capture chip.

[0062] Example 4

[0063] This embodiment provides an application of the single cell capture and transfer system described in Example 2 or the single cell analysis method described in Example 3 in drug screening, especially in high-throughput or low-throughput drug screening.

[0064] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A single cell capture and transfer system, characterized in that: The invention comprises a glass through-hole chip (2), a microgroove array chip (3) and a fluid capture chip, wherein the fluid capture chip comprises a horizontally arranged and flat-plate-shaped fluid capture chip body (1), the fluid capture chip body (1) having a cell suspension inlet (11) for adding a single cell suspension containing a plurality of single cells, and the lower end of the fluid capture chip body (1) has a plurality of spaced-apart single cell capture structures (12), and the plurality of single cell capture structures (12) are all connected to the cell suspension inlet (11), and the single cell capture structures (12) are used to capture any single cell in the single cell suspension; the fluid capture chip, the glass through-hole chip (2) and the microgroove array chip (3) are horizontally arranged in sequence from top to bottom and The glass through-hole chip (2) is overlapped with each other, and has a plurality of micropores (21) penetrating the glass through-hole chip (2) from top to bottom; the microgroove array chip (3) is concavely provided with a plurality of microgrooves (31), and the plurality of micropores (21), the plurality of microgrooves (31) and the plurality of single-cell capture structures (12) correspond to each other one by one, and each of the micropores (21) and the corresponding microgroove (31) are aligned with each other. Under the action of external force, the fluid capture chip can be horizontally moved at the upper end of the glass through-hole chip (2) to the point where the single-cell capture structure (12) thereon and the corresponding micropore (21) are staggeredly distributed to capture a single cell, or moved to align with each other to discharge the captured single cell through the corresponding micropore (21) into the corresponding microgroove (31).

2. The single cell capture and transfer system according to claim 1, characterized in that: The fluid capture chip body (1) further comprises a culture medium inlet (13) and a sample outlet (14) which are horizontally spaced apart, and the culture medium inlet (13), the cell suspension inlet (11) and the sample outlet (14) all penetrate the fluid capture chip body (1) from top to bottom, and the cell suspension inlet (11) is close to the culture medium inlet (13), and a microfluidic channel (15) is etched at the lower end of the fluid capture chip body (1), and the two ends of the microfluidic channel (15) are respectively connected to the culture medium inlet (13) and the sample outlet (14). The outlet (14) is connected, and the cell suspension inlet (11) is connected to one end of the microfluidic channel (15) close to the culture medium inlet (13), and multiple single cell capture structures (12) are etched on the lower end of the fluid capture chip body (1), and the single cell capture structure (12) has a liquid inlet (121) and a liquid outlet (122). Multiple single cell capture structures (12) are connected in series on the microfluidic channel (15) and are located between the cell suspension inlet (11) and the sample outlet (14).

3. The single cell capture and transfer system according to claim 2, characterized in that: The microfluidic channels (15) are provided in plurality, and a portion of each microfluidic channel (15) located between the cell suspension inlet (11) and the sample outlet (14) is respectively connected in series with a plurality of single cell capture structures (12).

4. The single cell capture and transfer system according to claim 2 or 3, characterized in that: The plurality of single cell capture structures (12) are distributed in an array at the lower end of the fluid capture chip body (1).

5. The single cell capture and transfer system according to claim 4, characterized in that: The single cell capture structure (12) is a groove horizontally etched at the lower end of the fluid capture chip body (1) and having a "U"-shaped horizontal cross-section. The two ends of the single cell capture structure are respectively its liquid inlet (121) and liquid outlet (122), and the middle part of the structure is a bend (123). A trumpet-shaped notch (124) is provided between the liquid inlet (121) and the bend (123), and the thin end of the notch is connected to the area between the bend (123) and the liquid outlet (122), and the notch (124) is used to capture single cells.

6. The single cell capture and transfer system according to claim 2, characterized in that: The upper surface of the microgroove array chip (3) is provided with a hydrophobic layer.

7. The single cell capture and transfer system according to claim 6, characterized in that: The hydrophobic layer is a super hydrophobic film.

8. A single cell analysis method using the single cell capture and transfer system according to any one of claims 2 to 7, characterized in that: The steps include: Step 1: taking a fluid capture chip and a through-glass chip (2), and stacking the fluid capture chip on the through-glass chip (2) in an offset manner; Step 2: adding culture medium to the culture medium inlet (13) of the fluid capture chip, and then taking a cell suspension using a capillary and inserting it into the cell suspension inlet (11); the cell suspension in the capillary flows along with the culture medium in the microfluidic channel (15) toward the sample outlet (14), and the single cell in the culture medium is captured by the single cell capture structure (12); Step 3: When the culture medium flows to the sample outlet (14), the microgroove array chip (3) is taken and stacked on the lower end of the through-glass chip (2), and each of the microholes (21) of the through-glass chip (2) is aligned with the corresponding microgroove (31) on the microgroove array chip (3); Step 4: horizontally slide the fluid capture chip until its single cell capture structure (12) is aligned with the corresponding micropores (21), so as to transfer the single cell captured by the single cell capture structure (12) to the corresponding microgroove (31) through the corresponding micropore (21); Step 5: After the single cell transfer in step 4 is completed, the microgroove array chip (3) is placed in a culture dish for single cell culture.

9. An application of the single cell capture and transfer system according to any one of claims 1 to 7 or the single cell analysis method according to claim 8 in drug screening.

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