Single cell sorting device

Abstract

The utility model relates to a single cell sorting device, and relates to the technical field of cell culture, the single cell sorting device comprises a conical tube, a microwell plate and electrodes, the microwell plate is connected to the end part of the conical tube, the electrodes comprise an inner electrode and an outer electrode which are respectively arranged inside and outside the conical tube, and the conical tube and the microwell plate are made of non-conductive materials and comprise plastic, glass and rubber. The inner electrode is fixed on the inner wall of the conical tube, the outer electrode is fixed on the outer wall of the conical tube, the distance between the end part of the inner electrode and the microwell plate is 1-3mm, and the end part of the outer electrode is flush with the microwell plate. The device provided by the utility model is simple in structure, realizes flexible distribution of the single cells by combining with a flow control and control system, realizes accurate distribution of the single cells, and reduces the influence on the cell activity at the same time.

Description

Technical Field

[0001] This application relates to the field of cell culture technology, and in particular to a single-cell sorting device. Background Technology

[0002] Single-cell analysis is a key focus of current medical diagnosis. Cells are heterogeneous, and traditional cell analysis techniques neglect the uniqueness of single cells. Therefore, single-cell analysis has important research significance. Single-cell capture is the first key step in single-cell analysis. Existing single-cell distribution methods include: (1) Limiting dilution method, which dilutes the cell population suspension with liquid to reduce its cell concentration, and then quantitatively distributes the diluted suspension to each culture dish. The probability of a single cell appearing in each culture dish is low. This method has a random component and cannot guarantee that there is only one cell in each culture dish, so the efficiency is low. (2) Flow cytometry distribution method, which requires fluorescent labeling and laser irradiation to excite fluorescence, which has an adverse effect on cell activity. (3) Image recognition method, Chinese patents CN118652743A and CN118325717A adopt the following method: first, prepare a cell suspension, then spray a tiny droplet with a spraying device, and the image recognition system determines whether there is a single cell in the droplet. If there is a single cell, the droplet is sent to the culture dish. If there is a droplet with 0 or more cells, it is sent to the waste liquid pool. This method is complex and requires microscope photography and an image recognition system.

[0003] Therefore, there is an urgent need for a single-cell sorting device that is simple in structure and highly accurate. Utility Model Content

[0004] The purpose of this application is to provide a single-cell sorting device with a simple structure that combines flow control and a control system to achieve flexible single-cell dispensing, thereby solving the inaccuracies and impacts on cell viability of existing dispensing methods.

[0005] The single-cell sorting device provided in this application adopts the following technical solution:

[0006] A single-cell sorting device includes a conical tube, a microplate, and electrodes. The microplate is connected to the end of the conical tube, and the electrodes include an inner electrode and an outer electrode, which are respectively disposed inside and outside the conical tube.

[0007] As a preferred technical solution of this application, the tapered tube and the microporous plate are made of non-conductive materials, including plastic, glass, and rubber.

[0008] As a preferred technical solution of this application, micropores are provided at the center of the microporous plate, the thickness of the microporous plate is set to 5-1000 micrometers, and the diameter of the micropores is set to 5-100 micrometers.

[0009] As a preferred technical solution of this application, the thickness of the microplate around the micropores is set to 10-100 micrometers; the diameter of the micropores is set to 10-40 micrometers.

[0010] As a preferred technical solution of this application, the tapered tube and the microporous plate are integrally formed or the microporous plate is sealed and connected to the narrow end of the tapered tube.

[0011] As a preferred technical solution of this application, the inner electrode is fixed on the inner wall of the conical tube, the outer electrode is fixed on the outer wall of the conical tube, the distance between the end of the inner electrode and the microporous plate is 1-3mm, and the end of the outer electrode is flush with the microporous plate.

[0012] As a preferred technical solution of this application, it also includes a culture dish, with the inner electrode fixed on the inner wall of the conical tube and the outer electrode fixed inside the culture dish.

[0013] In summary, this application includes at least one of the following beneficial technical effects:

[0014] 1. The sorting device of this application has a simple structure and is easy to operate, which can improve the sorting efficiency of single cells.

[0015] 2. This application utilizes micropores to dispense cells, thereby improving the accuracy of cell dispensing.

[0016] 3. By combining flow control and control systems, flexible single-cell distribution is achieved, reducing the impact on cell viability. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the device in Embodiment 1 of this application;

[0018] Figure 2 This is a schematic diagram of the device in Embodiment 2 of this application;

[0019] In the figure, 1 is a conical tube; 2 is an internal electrode; 3 is an external electrode; 4 is a microplate; and 5 is a petri dish. Detailed Implementation

[0020] The following is in conjunction with the appendix Figure 1 and 2 This application will be described in further detail below.

[0021] Example 1: This example proposes a single-cell sorting device, referring to... Figure 1 The device includes a tapered tube 1, a microporous plate 4, and electrodes.

[0022] The upper opening of the tapered tube 1 is larger than the lower opening. The microporous plate 4 is fixedly installed at the lower end of the tapered tube 1. The tapered tube 1 is made of a non-conductive material, including but not limited to plastic, glass, and rubber. In order to facilitate observation of the internal condition of the tapered tube 1, the tapered tube 1 in this embodiment can be a transparent PP tube, and the smaller diameter end is ultrasonically welded with a PP film as the microporous plate 4.

[0023] The microporous plate 4 is also made of a non-conductive material, including but not limited to plastic, glass, and rubber. The material can be the same as or different from that of the tapered tube 1. The tapered tube 1 and the microporous plate 3 can be integrally formed, or the microporous plate 4 can be sealed and connected to the narrow end of the tapered tube 1. When integrally formed, the same material is used. When sealed, the connection method can be bonding, welding, or other sealing methods.

[0024] The thickness of the microporous plate 4 is set to 5-1000 micrometers, preferably 10-100 micrometers at the center, and 20 micrometers in this embodiment. Microholes are formed at the center of the microporous plate 4, with diameters set to 5-100 micrometers, preferably 10-40 micrometers, and 30 micrometers in this embodiment. The methods for creating the microholes include, but are not limited to, laser drilling, mold-integrated hole forming, and microneedle puncture; laser drilling can be used in this embodiment.

[0025] The electrode includes an inner electrode 2 and an outer electrode 3. The inner electrode 2 is disposed inside the conical tube 1, and the outer electrode 3 is disposed outside the conical tube 1. In this embodiment, the inner electrode 2 is fixed to the inner wall of the conical tube 1. Specifically, the inner electrode 2 is bonded to the inner wall of the conical tube 1 using 316 stainless steel wire, and the distance between the lower end of the inner electrode 2 and the microporous plate 4 is 2 mm. The outer electrode 3 is fixed to the outer wall of the conical tube 1. Specifically, the outer electrode 3 is bonded to the outer wall of the conical tube 1 using 316 stainless steel wire, and the lower end of the outer electrode 3 is flush with the lower surface of the microporous plate 4.

[0026] The electrodes can be made of various conductive materials, with stainless steel or alloys having good biocompatibility being preferred. The electrode structure includes, but is not limited to, conductive wires, sprayed conductive materials, and bonded conductive sheets.

[0027] It should be noted that the basic principle of signal generation when cells pass through is the Coulter principle. The cell culture medium is conductive, and the cells are poor conductors of electricity. When the device of this application enters the culture container for dispensing, the two electrodes inside and outside the conical tube 1 conduct electricity through the cell culture medium. When the cells pass through the micropores, the conductive channel is temporarily blocked, so the resistance of the two electrodes changes abruptly, and the control system can sense this signal.

[0028] Example 2: This example proposes a single-cell sorting device, referring to... Figure 2The device includes a conical tube 1, a microplate 4, electrodes, and a culture dish 5. The difference between this embodiment and embodiment 1 is that the fixing position of the outer electrode 3 is different. In this embodiment, the inner electrode 2 is fixed on the inner wall of the conical tube 1, and the outer electrode 3 is fixed in the culture dish 5 and is not connected to the conical tube 1.

[0029] The embodiments described in this specific implementation are preferred embodiments of this application and are not intended to limit the scope of protection of this application. Identical components are represented by the same reference numerals. Therefore, all equivalent changes made to the structure, shape, and principle of this application should be covered within the scope of protection of this application.

Claims

1. A single cell sorting device, characterized by, The application relates to a microplate electrode, which comprises a conical tube (1), a microplate (4) and electrodes, the microplate (4) is connected to the end of the conical tube (1), and the electrodes comprise an inner electrode (2) and an outer electrode (3) which are arranged in the inner and outer parts of the conical tube (1) respectively.

2. The single cell sorting device of claim 1, wherein, The conical tube (1) and the microplate (4) are made of non-conductive materials, such as plastic, glass and rubber.

3. The single cell sorting device of claim 1, wherein, The microplate (4) is provided with a microhole in the center, the thickness of the microplate (4) is 5-1000 microns, and the diameter of the microhole is 5-100 microns.

4. A single cell sorting device according to claim 3, wherein, The thickness of the microplate (4) around the microhole is 10-100 microns, and the diameter of the microhole is 10-40 microns.

5. The single cell sorting device of claim 1, wherein, The conical tube (1) and the microplate (4) are integrally formed or the microplate (4) is sealingly connected to the thin end of the conical tube (1).

6. A single cell sorting device according to any one of claims 1 to 5, wherein, The inner electrode (2) is fixed to the inner wall of the conical tube (1), the outer electrode (3) is fixed to the outer wall of the conical tube (1), the end of the inner electrode (2) is 1-3 mm away from the microplate (4), and the end of the outer electrode (3) is flush with the microplate (4).

7. A single cell sorting device according to any one of claims 1 to 5, wherein, The application further discloses a culture dish (5), the inner electrode (2) is fixed to the inner wall of the conical tube (1), and the outer electrode (3) is fixed to the culture dish (5).

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