Combined micro-fluidic chip for separating and enriching circulating tumor cells

Abstract

The utility model relates to the technical field of micro-fluidic chips, and discloses a combined micro-fluidic chip for separating and enriching circulating tumor cells, which comprises a chip main body, the chip main body comprises a first chip and a second chip, the first chip is provided with a charge trapping liquid cavity, a reaction bin, a sample inlet and a sample outlet, and the second chip is provided with a charge trapping liquid outlet. The sample inlet and the sample outlet are communicated through a connecting flow channel, and a magnet is arranged below the connecting flow channel; a tip placing area, a confining liquid pool, a first coloring agent pool, a second coloring agent pool, a stationary liquid pool, a permeabilizing agent pool and a cell filter are arranged on the second chip. The utility model provides a novel method for detecting and separating CTC based on surface charges of cancer cells as pre-capture of the CTC, and then the CTC is integrally captured by adopting a physical and immune double-enrichment system, so that the capture rate of the CTC is greatly improved, and technical support is provided for subsequent automatic and batch detection; the filtering efficiency and the detection precision are greatly improved.

Description

Technical Field

[0001] The utility model relates to the technical field of microfluidic chips, in particular to a combined microfluidic chip for separating and enriching circulating tumor cells. Background Art

[0002] Circulating tumor cells (CTCs) are tumor cells that spontaneously shed from the main lesions. They are present in very small amounts in peripheral blood, with only one or a few cells in every 10 ml of blood. However, they also contain about 100 million white blood cells and about 50 billion red blood cells. Therefore, the sensitivity and specificity of their sorting are also required to be high, and early detection and characterization of CTCs are crucial for the diagnosis and proper treatment of cancer. In the prior art, the detection of circulating tumor cells is mainly divided into the separation, enrichment, and identification and analysis of circulating tumor cells. In the prior art, due to the small number of CTCs, clinical testing generally relies on drawing 10-20 ml of blood for clinical testing, which may lead to missed diagnosis. Utility Model Content

[0003] The utility model aims to solve the problems existing in the prior art and proposes a combined microfluidic chip for separating and enriching circulating tumor cells.

[0004] In order to achieve the above purpose, the utility model adopts the following technical solutions:

[0005] A combined microfluidic chip for separating and enriching circulating tumor cells comprises a chip body, wherein the chip body comprises a first chip and a second chip, wherein the first chip is provided with a charge capture liquid cavity, a reaction chamber, an inlet and an outlet, wherein the inlet and the outlet are connected via a connecting flow channel, and a magnet is provided below the connecting flow channel; and the second chip is provided with a tip placement area, a closed liquid pool, a first dye pool, a second dye pool, a fixative pool, a permeabilizing agent pool and a cell filter.

[0006] Preferably, a cleaning liquid pool is disposed on both the first chip and the second chip, and a waste liquid pool is disposed on the first chip.

[0007] Preferably, a tip head is arranged inside the tip head placement area, and the size of the tip head is adapted to the opening size of the charge capture liquid cavity, the reaction chamber, the sample inlet and the sample outlet.

[0008] Preferably, the connecting flow channel is serpentine.

[0009] Preferably, the connecting channel contains a protrusion structure for adsorbing CTCs in the sample thereon by electric charge.

[0010] Preferably, both ends of the connecting flow channel are connected to the bottom of the sample inlet and the sample outlet respectively.

[0011] Preferably, the first chip is in the shape of a box, the charge capture liquid chamber and the reaction chamber are arranged at a corner of the first chip, the cleaning liquid pool and the waste liquid pool in the first chip are arranged on one side of the charge capture liquid chamber, the sample inlet and the sample outlet are arranged in parallel with the cleaning liquid pool and the waste liquid pool, the tip head placement area and the cell filter are respectively located at the two ends of the second chip, the closing liquid pool, the first stain pool, the second stain pool, the fixative liquid pool, and the permeabilizing agent pool are arranged between the tip head placement area and the cell filter, and the closing liquid pool, the first stain pool, the second stain pool, the fixative liquid pool, and the permeabilizing agent pool are in a rectangular array, and the cleaning liquid pool on the second chip is located between the fixative liquid pool and the permeabilizing agent pool.

[0012] Compared with the prior art, the beneficial effects of the utility model are:

[0013] The utility model proposes a new CTC detection and separation method based on the surface charge of cancer cells as the pre-capture of CTCs, and then uses a physical + immune dual enrichment system to capture CTCs in an integrated manner, which greatly improves the capture rate of CTCs and provides technical support for subsequent automated and batch detection, greatly improving the filtration efficiency and detection accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is a three-dimensional schematic diagram of the composition of a combined microfluidic chip for separating and enriching circulating tumor cells proposed by the utility model;

[0015] Figure 2 This is a top view of a combined microfluidic chip for separating and enriching circulating tumor cells proposed by the utility model.

[0016] In the figure: 1. Charge capture liquid chamber; 2. Reaction chamber; 3. Sample inlet; 4. Magnet; 5. Connecting channel; 6. Sample outlet; 7. Cleaning liquid pool; 8. Waste liquid pool; 9. Tip head placement area; 10. Sealing liquid pool; 11. First stain pool; 12. Second stain pool; 13. Fixative pool; 14. Permeabilization agent pool; 15. Cell filter; 16. First chip; 17. Second chip. DETAILED DESCRIPTION

[0017] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments.

[0018] Reference Figure 1 and Figure 2A combined microfluidic chip for separating and enriching circulating tumor cells comprises a chip body, wherein the chip body comprises a first chip 16 and a second chip 17, wherein the first chip 16 is provided with a charge capture liquid chamber 1, a reaction chamber 2, an inlet 3 and an outlet 6, wherein the inlet 3 and the outlet 6 are connected via a connecting channel 5, and a magnet 4 is provided below the connecting channel 5; the second chip 17 is provided with a tip head placement area 9, a closing liquid pool 10, a first dye pool 11, a second dye pool 12, a fixing liquid pool 13, a permeabilizing agent pool 14 and a cell filter 15, wherein the tip head placement area 9 is used for placing a tip head, the closing liquid pool 10 is used for storing closing liquid, the first dye pool 11 is used for storing a first dye, the second dye pool 12 is used for storing a second dye, the fixing liquid pool 13 is used for storing a fixing liquid, and the permeabilizing agent pool 14 is used for storing a permeabilizing agent, and the first dye and the second dye can be selected according to actual conditions.

[0019] When the device is in use, first add the sample solution and the charge capture solution (in the charge capture liquid chamber 1) to the reaction chamber 2, add them to the injection port 3 after the reaction is completed, apply positive and negative pressure to the channel (realized by the pressure gun head) to make the solution completely adsorbed in the connecting channel 5 (through the action of the magnet 4 below), and impurities will also be attached, and then use the pipette head to add cleaning liquid to the inlet to clean the impurities in the connecting channel 5 (add the pressure gun head to the injection port 3, and make the test liquid flow repeatedly in the connecting channel 5 through positive and negative pressure), remove the waste liquid through the pipette, then remove the bottom magnet 4, add cleaning liquid, release the CTC cells and aggregate them through the magnet 4 at the outlet, and then put the first chip The CTC solution purified by charge capture in 16 is transferred into the cell filter 16, and the fixing solution → cleaning solution → permeabilizing solution → cleaning solution → blocking solution → cleaning solution → first stain → cleaning solution → second stain → cleaning solution are added to the cell filter 16 in sequence. After sufficient reaction, it can be detected. Compared with the existing technology, this device proposes a new CTC detection and separation method based on the surface charge of cancer cells as the pre-capture of CTC, and then uses a physical + immune double enrichment system to capture CTC in an integrated manner, which greatly improves the capture rate of CTC and provides technical support for subsequent automated and batch detection, greatly improving the filtration efficiency and detection accuracy.

[0020] In this embodiment, a cleaning liquid pool 7 is disposed on both the first chip 16 and the second chip 17. The cleaning liquid pool 7 is used to store cleaning liquid. A waste liquid pool 8 is disposed on the first chip 16 to facilitate moving waste liquid into the waste liquid pool 8 by a pipette.

[0021] In this embodiment, a tip head is arranged inside the tip head placement area 9, and the reagent part is added into the corresponding position in sequence through the machine-operated tip head. The size of the tip head is adapted to the opening size of the charge capture liquid chamber 1, the reaction chamber 2, the sample inlet 3, and the sample outlet 6, so as to facilitate sample addition and sampling.

[0022] In this embodiment, the connecting channel 5 is serpentine, which can increase the contact area between the connecting channel 5 and the magnet 4 .

[0023] In this embodiment, the connecting channel 5 contains a protrusion structure for adsorbing CTCs in the sample thereon through electric charge.

[0024] In this embodiment, two ends of the connecting flow channel 5 are connected to the bottom of the sample inlet 3 and the sample outlet 6 respectively.

[0025] In this embodiment, the first chip 16 is in the shape of a box, the charge capture liquid chamber 1 and the reaction chamber 2 are arranged at a corner of the first chip 16, the cleaning liquid pool 7 and the waste liquid pool 8 in the first chip 16 are arranged on one side of the charge capture liquid chamber 1, the sample inlet 3 and the sample outlet 6 are arranged in parallel with the cleaning liquid pool 7 and the waste liquid pool 8, the tip head placement area 9 and the cell filter 15 are respectively located at the two ends of the second chip 17, the closing liquid pool 10, the first stain pool 11, the second stain pool 12, the fixing liquid pool 13, and the permeabilizing agent pool 14 are arranged between the tip head placement area 9 and the cell filter 15, and the closing liquid pool 10, the first stain pool 11, the second stain pool 12, the fixing liquid pool 13, and the permeabilizing agent pool 14 are in a rectangular array, and the cleaning liquid pool 7 on the second chip 17 is located between the fixing liquid pool 13 and the permeabilizing agent pool 14. Through reasonable space arrangement, the volume of the device can be made more compact and streamlined.

[0026] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes within the technical scope disclosed by the present invention according to the technical scheme and the utility model concept of the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A combined microfluidic chip for separating and enriching circulating tumor cells, comprising a chip body, characterized in that: The chip body comprises a first chip (16) and a second chip (17); the first chip (16) is provided with a charge capture liquid chamber (1), a reaction chamber (2), an injection port (3) and an output port (6); the injection port (3) and the output port (6) are connected via a connecting flow channel (5); a magnet (4) is provided below the connecting flow channel (5); the second chip (17) is provided with a tip placement area (9), a closed liquid pool (10), a first dye pool (11), a second dye pool (12), a fixative liquid pool (13), a permeabilizing agent pool (14) and a cell filter (15).

2. The combined microfluidic chip for separating and enriching circulating tumor cells according to claim 1, characterized in that: A cleaning liquid pool (7) is provided on both the first chip (16) and the second chip (17), and a waste liquid pool (8) is provided on the first chip (16).

3. The combined microfluidic chip for separating and enriching circulating tumor cells according to claim 1, characterized in that: A tip head is arranged inside the tip head placement area (9), and the size of the tip head is adapted to the opening sizes of the charge capture liquid chamber (1), the reaction chamber (2), the sample inlet (3), and the sample outlet (6).

4. The combined microfluidic chip for separating and enriching circulating tumor cells according to claim 1, characterized in that: The connecting flow channel (5) is in a serpentine shape.

5. The combined microfluidic chip for separating and enriching circulating tumor cells according to claim 4, characterized in that: The connecting flow channel (5) contains a protrusion structure inside, which is used to adsorb CTCs in the sample on it through electric charge.

6. The combined microfluidic chip for separating and enriching circulating tumor cells according to claim 1, characterized in that: The two ends of the connecting flow channel (5) are respectively connected to the bottom of the sample inlet (3) and the sample outlet (6).

7. The combined microfluidic chip for separating and enriching circulating tumor cells according to claim 2, characterized in that: The first chip (16) is in the shape of a box body, the charge capture liquid chamber (1) and the reaction chamber (2) are arranged at a corner of the first chip (16), the cleaning liquid pool (7) and the waste liquid pool (8) in the first chip (16) are arranged on one side of the charge capture liquid chamber (1), the sample inlet (3) and the sample outlet (6) are arranged in parallel with the cleaning liquid pool (7) and the waste liquid pool (8), the tip head placement area (9) and the cell filter (15) are respectively located at two ends of the second chip (17), and the closed The liquid pool (10), the first stain pool (11), the second stain pool (12), the fixative liquid pool (13), and the permeabilizing agent pool (14) are arranged between the tip head placement area (9) and the cell filter (15), and the closed liquid pool (10), the first stain pool (11), the second stain pool (12), the fixative liquid pool (13), and the permeabilizing agent pool (14) are in a rectangular array shape, and the cleaning liquid pool (7) on the second chip (17) is located between the fixative liquid pool (13) and the permeabilizing agent pool (14).

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