Reusable single cell capturing and releasing electrochemical analysis micro-fluidic chip and method

Abstract

The invention discloses an electrochemical analysis micro-fluidic chip capable of repeatedly capturing and releasing single cells and a method, particularly relates to the technical field of single cell analysis and microfluidics, and solves the problem that in the prior art, captured cells cannot be released after single cells are captured and detected by a single cell analysis micro-fluidic technology. The technical problems that a new micro-fluidic chip needs to be manufactured again when repeated experiments are carried out, so that materials are consumed, the cost is increased, and the detection efficiency is reduced due to the fact that time is needed for remanufacturing are solved. According to the technical scheme, a micro-fluidic technology is combined with electrochemical detection, an integral structure of a micro-fluidic chip is designed, and a single cell can be released after the captured single cell is detected; the chip can be repeatedly used and has high integration, the time for repeatedly manufacturing the chip is greatly shortened, and the cost is further saved.

Description

Technical Field

[0001] The present invention relates to the field of single cell analysis and microfluidics, and in particular to a reusable microfluidics chip and method for electrochemical analysis of captured and released single cells. Background Art

[0002] Traditional single-cell analysis methods have some problems, such as cumbersome operation process, expensive equipment or difficulty in maintaining cell activity. The existing single-cell analysis microfluidic technology cannot release the captured cells after capturing and detecting single cells. When repeating the experiment, new microfluidic chips must be re-manufactured, which not only consumes materials and increases costs, but also reduces detection efficiency due to the time required for re-manufacturing. Summary of the invention

[0003] Therefore, the present invention solves the technical problem that the single-cell analysis microfluidic technology in the prior art cannot release the captured cells after capturing and detecting the single cells, and a new microfluidic chip must be remanufactured when repeating the experiment, which not only consumes materials and increases costs, but also reduces the detection efficiency due to the time required for remanufacturing. The present invention combines microfluidic technology with electrochemical detection to design an overall structure of a microfluidic chip, which can release the captured single cells after detecting the captured single cells, solving the problem that a new microfluidic chip must be remanufactured when repeating the experiment, which allows the chip to be reused, while greatly reducing the time for repeated chip manufacturing, improving detection efficiency, and further saving costs.

[0004] To achieve the above-mentioned purpose, the present invention provides a reusable microfluidic chip for capturing and releasing single cells for electrochemical analysis, comprising a glass substrate on which a polydimethylsiloxane layer is disposed; an electrode is disposed on one side of the glass substrate bonded to the polydimethylsiloxane layer; and a chip is disposed inside the polydimethylsiloxane layer.

[0005] Furthermore, the polydimethylsiloxane layer is provided with an inlet 1 and an outlet 1, and the polydimethylsiloxane layer is provided with a capture chamber for connection; the two sides of the capture chamber are respectively connected with the inlet 1 and the outlet 1 through a channel 1.

[0006] Furthermore, the polydimethylsiloxane layer is provided with an inlet 2 and an outlet 2, the inlet 2 and the outlet 2 are connected to a channel 2 provided inside the polydimethylsiloxane layer, and the channel 2 is connected to the capture chamber via a channel 3. The channel 1 and the channel 2 are arranged in parallel.

[0007] Furthermore, there are multiple capture chambers in a horizontal array, and the capture chambers are tilted toward the outlet to facilitate the flow of the solution; the capture chambers on both sides are connected to channel 1. Each capture chamber has the same size, and the capture of cells of different sizes can be achieved by adjusting the size of the capture chamber.

[0008] Furthermore, each of the capture chambers is respectively connected to channel three.

[0009] Furthermore, the width of the channel three is smaller than the diameter of the captured cells, thereby achieving the capture, release and electrochemical detection of cells of different sizes.

[0010] Furthermore, an electrode is disposed under each capture chamber, and the electrode is connected to an electrochemical workstation, so that the captured cells can be electrochemically detected.

[0011] Furthermore, the electrode is connected to an electrochemical workstation.

[0012] Furthermore, the inlet 1, the inlet 2, the outlet 1 and the outlet 2 penetrate the polydimethylsiloxane layer.

[0013] The present invention also provides a method for using the above-mentioned reusable capture and release single cell electrochemical analysis microfluidic chip, comprising the following steps:

[0014] S1: The solution of cells to be detected is pumped into the inlet 1, and the cell solution enters the capture chamber through the channel 1. When flowing through the capture chamber, the cells can be captured in the capture chamber, and most of the excess liquid flows out from the outlet 1 through the channel 1, and a small amount of liquid flows into the channel 2 through the channel 3;

[0015] S2: After the cell capture is completed, the electrode of the electrochemical workstation can be connected to the electrode arranged below the capture chamber to perform electrochemical detection on the cells in the capture chamber; if it is necessary to observe and detect the cells for a long time, the cell culture medium can be pumped into the inlet 1 to keep the cells active;

[0016] S3: After the electrochemical detection is completed, pure water is pumped into inlet two, part of the pure water flows through channel two and flows out from outlet two, and part of the pure water flows from channel two through channel three to the capture chamber, releasing the captured cells, and the released cells flow out from outlet one through channel one.

[0017] In the above technical solution, the technical effects and advantages provided by the present invention are:

[0018] 1. The reusable capture and release single cell electrochemical analysis microfluidic chip and method provided by the present invention can realize the capture and release and electrochemical detection of cells of different sizes by adjusting the size and width of the capture chamber and channel three, and the width of channel three needs to be smaller than the diameter of the captured cell. By changing the number of capture chamber arrays, different experimental conditions can be met.

[0019] 2. The reusable capture and release single-cell electrochemical analysis microfluidic chip and method provided by the present invention combine cell capture and release with electrochemical detection through the use of microfluidic technology, and can realize single-cell capture and release and detection on a chip. At the same time, the chip can be reused and has high integration. At the same time, it greatly reduces the time for repeated chip production, further saving costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For ordinary technicians in this field, other drawings can also be obtained based on these drawings.

[0021] Figure 1 This is a schematic diagram of the overall structure of the reusable microfluidic chip for capturing and releasing single cells for electrochemical analysis of the present invention;

[0022] Figure 2 A top view of the microfluidic chip for reusable capture and release of single cells for electrochemical analysis according to the present invention.

[0023] Description of reference numerals:

[0024] 1. Inlet one; 2. Channel one; 3. Capture chamber; 4. Outlet one; 5. Inlet two; 6. Channel three; 7. Channel two; 8. Outlet two; 9. Glass substrate; 10. Dimethylsiloxane layer; 11. Electrode. DETAILED DESCRIPTION

[0025] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.

[0026] Embodiment 1:

[0027] This embodiment provides a reusable microfluidic chip for capturing and releasing single cells for electrochemical analysis, including a glass substrate 9, on which a polydimethylsiloxane layer 10 is disposed; an electrode 11 is disposed on one side of the glass substrate 9 bonded to the polydimethylsiloxane layer 10; and a chip is disposed inside the polydimethylsiloxane layer 10. The polydimethylsiloxane layer 10 is provided with an inlet 1 and an outlet 4, and the polydimethylsiloxane layer 10 is provided with a capture chamber 3 for connection; the capture chamber 3 is connected to the inlet 1 and the outlet 4 on both sides through a channel 2, respectively. The polydimethylsiloxane layer 10 is provided with an inlet 2 5 and an outlet 2 8, and the inlet 2 5 and the outlet 2 8 are connected to the channel 2 7 disposed inside the polydimethylsiloxane layer 10, and the channel 2 7 is connected to the capture chamber 3 through a channel 3 6. There are multiple capture chambers 3 in a horizontal array, and the capture chambers 3 on both sides are connected to the channel 2.

[0028] Each of the capture chambers 3 is connected to a channel 3 6. The width of the channel 3 6 is smaller than the diameter of the captured cell. An electrode 11 is disposed below each of the capture chambers 3. The electrode 11 is connected to an electrochemical workstation. The inlet 1, the inlet 2 5, the outlet 1 4 and the outlet 2 8 penetrate the polydimethylsiloxane layer 10.

[0029] A solution of cells to be detected is pumped in from inlet 1, and the cell solution enters the capture chamber 3 through channel 1 2. When flowing through the capture chamber 3, the cells can be captured in the capture chamber 3, and most of the excess liquid flows out from outlet 1 4 through channel 1 2, and a small amount of liquid flows into channel 2 7 through channel 3 6. After the cell capture is completed, the electrode 11 of the electrochemical workstation can be connected to the electrode 11 set below the capture chamber 3 to perform electrochemical detection on the cells in the capture chamber 3. If it is necessary to observe and detect the cells for a long time, the cell culture fluid can be pumped in from inlet 1 to keep the cells active. After the electrochemical detection is completed, pure water is pumped into inlet 2 5, and part of the pure water flows out from outlet 2 8 through channel 2 7, and part of the pure water flows from channel 2 7 through channel 3 6 to the capture chamber 3, releasing the captured cells, and the released cells flow out from outlet 4 through channel 1 2. By adjusting the size and width of the capture chamber 3 and channel 3 6, and at the same time, the width of channel 3 6 needs to be smaller than the diameter of the captured cells, the capture, release and electrochemical detection of cells of different sizes can be achieved. By changing the number of capture chamber 3 arrays, different experimental conditions can be met. By using microfluidics technology to combine cell capture and release with electrochemical detection, single cell capture and release and detection can be achieved on a chip. At the same time, the chip can be reused, with high integration, and the time for repeated chip production can be greatly reduced, further saving costs.

[0030] Embodiment 2:

[0031] This embodiment provides a method for using the above-mentioned reusable capture and release single cell electrochemical analysis microfluidic chip, comprising the following steps:

[0032] S1: The cell solution to be detected is pumped into the inlet 1, and the cell solution enters the capture chamber 3 through the channel 2. When flowing through the capture chamber 3, the cells can be captured in the capture chamber 3, and most of the excess liquid flows out from the outlet 4 through the channel 2, and a small amount of liquid flows into the channel 2 7 through the channel 3 6;

[0033] S2: After the cell capture is completed, the electrode 11 of the electrochemical workstation can be connected to the electrode 11 disposed below the capture chamber 3 to perform electrochemical detection on the cells in the capture chamber 3; if it is necessary to observe and detect the cells for a long time, the cell culture fluid can be pumped into the inlet 1 to keep the cells active;

[0034] S3: After the electrochemical detection is completed, pure water is pumped into inlet 2 5, part of the pure water flows through channel 2 7 and flows out from outlet 2 8, and part of the pure water flows from channel 2 7 through channel 3 6 to the capture chamber 3, releasing the captured cells, and the released cells flow out from outlet 1 4 through channel 1 2.

[0035] The above description is only by way of illustration of certain exemplary embodiments of the present invention. It is undoubted that those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the above drawings and descriptions are illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.

Claims

1. A reusable microfluidic chip for capturing and releasing single cells for electrochemical analysis, characterized in that: The invention comprises a glass substrate (9), on which a polydimethylsiloxane layer (10) is arranged; an electrode (11) is arranged on one side of the glass substrate (9) that is bonded to the polydimethylsiloxane layer (10); and a chip is arranged inside the polydimethylsiloxane layer (10).

2. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 1, characterized in that: The chip comprises an inlet (1) and an outlet (4) opened on a polydimethylsiloxane layer (10), and a capture chamber (3) is opened inside the polydimethylsiloxane layer (10); the two sides of the capture chamber (3) are respectively connected to the inlet (1) and the outlet (4) through a channel (2).

3. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 2, characterized in that: The polydimethylsiloxane layer (10) is provided with an inlet 2 (5) and an outlet 2 (8), and the inlet 2 (5) and the outlet 2 (8) are respectively connected to a channel 2 (7) provided inside the polydimethylsiloxane layer (10), and the channel 2 (7) is connected to the capture chamber (3) via a channel 3 (6).

4. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 3, characterized in that: There are a plurality of capture chambers (3) arranged in a horizontal array, and the capture chambers (3) on both sides are connected to channel 1 (2).

5. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 4, characterized in that: Each of the capture chambers (3) is respectively connected to channel three (6).

6. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 5, characterized in that: The width of the channel three (6) is smaller than the diameter of the captured cells.

7. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 6, characterized in that: An electrode (11) is arranged below each capture chamber (3).

8. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 7, characterized in that: The electrode (11) is connected to an electrochemical workstation.

9. The reusable capture and release single cell electrochemical analysis microfluidic chip according to claim 8, characterized in that: The inlet 1 (1), the inlet 2 (5), the outlet 1 (4) and the outlet 2 (8) penetrate the polydimethylsiloxane layer (10).

10. A method for using the reusable capture and release single cell electrochemical analysis microfluidic chip according to any one of claims 1 to 9, characterized in that: The steps include: S1: A solution of cells to be detected is pumped into the inlet 1 (1), and the cell solution enters the capture chamber (3) through the channel 1 (2). When the solution flows through the capture chamber (3), the cells are captured in the capture chamber (3), and the excess liquid flows out from the outlet 1 (4) through the channel 1 (2), and part of the liquid flows into the channel 2 (7) through the channel 3 (6); S2: After the cell capture is completed, the electrode (11) of the electrochemical workstation is connected to the electrode (11) arranged below the capture chamber (3), and the cells in the capture chamber (3) are electrochemically detected; if it is necessary to observe the detected cells for a long time, the cell culture medium is pumped into the inlet 1 (1); S3: After the electrochemical detection is completed, pure water is pumped into inlet two (5), part of the pure water flows through channel two (7) and flows out from outlet two (8), and part of the pure water flows from channel two (7) through channel three (6) to the capture chamber (3), releasing the captured cells, and the released cells flow out from outlet one (4) through channel one (2).

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