Multi-channel cell separation device based on magnetic nanoparticles

Abstract

The utility model relates to the field of biomedical engineering, in particular to a multi-channel cell separation device based on magnetic nanoparticles, which comprises an incubator, a first input pipe and a second input pipe communicated with the incubator, a plurality of channel pipes communicated with the incubator, a temporary storage pipe communicated with each channel pipe, and a plurality of magnetic nanoparticles communicated with the temporary storage pipe. One end, far away from the channel pipe, of the temporary storage pipe is arranged upwards, a coil is arranged on the temporary storage pipe, one end, far away from the incubator, of the channel pipe is provided with a first control valve, one end, far away from the channel pipe, of the temporary storage pipe is provided with a second control valve, and the second control valve is communicated with a collecting chamber; and a valve is arranged between each channel pipe and the incubator. The cell separation device has the effects that the cell separation operation is simplified, and the cell separation efficiency is improved.

Description

Technical Field

[0001] The utility model relates to the field of biomedical engineering, in particular to a multi-channel cell separation device based on magnetic nanoparticles. Background Art

[0002] In biomedical research and clinical practice, isolating and analyzing specific cell types is a key step. Existing cell separation methods primarily include flow cytometry, density gradient centrifugation, and magnetic separation. Magnetic separation utilizes magnetic nanoparticles that bind to antibodies on the surface of target cells, achieving cell separation under the influence of a magnetic field.

[0003] Magnetic separation methods specifically include MACS (Magnetic-Activated Cell Sorting) technology, Dynabeads technology, EasySep technology and CellSearch technology.

[0004] MACS technology is a cell separation system based on superparamagnetic nanoparticles. It uses superparamagnetic beads, approximately 50 nanometers in diameter, coated with specific antibodies or ligands that bind to surface markers on target cells. Labeled cells are then separated using a MACS column placed in a strong magnetic field. Due to the small size of the magnetic beads, they do not cause mechanical damage to the cells, ensuring cell activity. However, this technology can only be applied to certain cell types, such as T cells and B cells.

[0005] Dynabeads technology also uses superparamagnetic beads, but the superparamagnetic beads have a larger diameter (usually between 1-4.5 microns) and can carry more antibodies or ligands. The labeled cells are then separated from the sample using a magnetic rack. Due to the relatively large diameter of the superparamagnetic beads, cells can be separated more quickly and efficiently under a magnetic field. However, it can only be applied to certain types of cells, and due to the increased diameter of the superparamagnetic beads, there is a possibility of affecting cell activity.

[0006] EasySep technology uses magnetic nanoparticles and an optimized immunoseparation procedure. The magnetic nanoparticles bind to target cells via specific antibodies. During separation, the sample tube is placed in a specially designed magnet, where target cells are attracted to the tube wall, while non-target cells are removed by decanting. This technology allows for efficient cell separation, but only one cell type can be isolated at a time.

[0007] CellSearch technology uses magnetic nanoparticles coated with antibodies to label CTCs in the blood. These labeled cells are then separated from the blood using a magnetic field and examined using a fluorescence microscope. While this method allows for efficient and accurate cell separation, it is expensive.

[0008] However, in the above method, when faced with a mixture of multiple cells, it is necessary to separate the multiple cells one by one. Each time the separation of one cell is completed, the above steps need to be repeated, which is a cumbersome operation. In addition, in the above method, there are many manual operations, which can easily lead to incomplete cell separation each time the cells are separated, resulting in varying degrees of cell loss. Utility Model Content

[0009] In order to solve the problems of complicated operation and large loss when separating multiple cells one by one, the utility model provides a multi-channel cell separation device based on magnetic nanoparticles.

[0010] The utility model provides a multi-channel cell separation device based on magnetic nanoparticles, which adopts the following technical solutions:

[0011] A multi-channel cell separation device based on magnetic nanoparticles includes an incubator, which is connected to a first input tube and a second input tube. The incubator is connected to multiple channel tubes, each of which is connected to a temporary storage tube, the temporary storage tube is arranged upward at one end away from the channel tube, a coil is provided on the temporary storage tube, a first control valve is provided at one end of the channel tube away from the incubator, a second control valve is provided at one end of the temporary storage tube away from the channel tube, the second control valve is connected to a collection chamber, and a valve is provided between each channel tube and the incubator.

[0012] In a specific embodiment, the first control valve is connected to the incubator through a pipeline.

[0013] In a specific embodiment, the temporary storage tube has an inner diameter of 1-3 cm and a length of 5-15 cm.

[0014] In a specific embodiment, the inner diameter of the coil is 1.5-3.5 cm, the number of turns of the coil is 20-60, and the interval between each turn is 1-3 mm.

[0015] In a specific embodiment, each of the collection chambers is provided with a counter.

[0016] In a specific embodiment, a delivery pump is connected between the incubator and the channel tube.

[0017] In a specific embodiment, a three-way valve is connected between the delivery pump and the incubator, and the three-way valve is connected to a buffer storage device.

[0018] In a specific embodiment, the incubator is connected to a cleaning tube.

[0019] In a specific embodiment, the incubator is connected to a waste liquid recovery tank.

[0020] In a specific possible implementation manner, the temporary storage tube is arranged vertically.

[0021] In summary, the present invention has at least one of the following beneficial technical effects:

[0022] 1. During cell separation, a valve controls one channel to connect to the incubator, while the other channels are closed, separating one cell type from the mixture. The mixture is then re-introduced into the incubator, where different antibodies and magnetic nanoparticles are added. The valve controls the connection of the remaining channels, closing the previously opened channels, and performing additional cell separations until all cells are separated. This eliminates the need to clean the channels after each cell separation, simplifying the cell separation process and improving cell separation efficiency.

[0023] 2. The magnetic field generated by the coil is used to separate the cells of the entire mixture stored in the incubator in the temporary storage tube, reducing manual participation and reducing cell loss during the cell separation process.

[0024] 3. A counter is installed in the collection chamber to facilitate automatic counting of separated cells and subsequent analysis of the separated cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a schematic diagram of the overall structure.

[0026] Figure 2 It is a cross-sectional view showing the channel tube structure.

[0027] Explanation of the accompanying drawings: 1. Incubator; 2. First input pipe; 3. Second input pipe; 4. Delivery pump; 5. Channel pipe; 6. Coil; 7. Collection chamber; 8. Valve; 9. Cleaning pipe; 10. Waste liquid recovery tank; 11. Temporary storage pipe; 12. First control valve; 13. Second control valve; 14. Three-way valve; 15. Buffer storage device. DETAILED DESCRIPTION

[0028] The following is combined with Figure 1-2 The utility model is described in further detail.

[0029] Reference Figure 1The multi-channel cell separation device based on magnetic nanoparticles includes a low-temperature cell magnetic nanoparticle incubator 1 (hereinafter referred to as the incubator) and a multi-channel fluid separation component. The incubator 1 is connected to a first input tube 2 for inputting a cell flow into the incubator 1, and the incubator 1 is connected to a second input tube 3 for transporting magnetic nanoparticles with different antibodies into the incubator 1. The outlet of the incubator 1 is connected to a three-way valve 14, one outlet of the three-way valve 14 is connected to a delivery pump 4, and the other outlet is connected to a buffer storage device 15, and the outlet of the delivery pump 4 is connected to the multi-channel fluid separation component. It can be understood that the cell flow is a mixture of cells and buffer, and the cells are driven by the flow of the buffer to achieve movement.

[0030] Reference Figure 1 The incubator 1 has temperature control and stirring functions. The cell stream is pre-input into the incubator 1 through the first input tube 2, and the magnetic nanoparticles mixed with antibodies are input into the incubator 1 through the second input tube 3. After the cell stream, the mixture of antibodies with different magnetic nanoparticles, or a specific single antibody with magnetic nanoparticles are input into the barrel 11, the incubator 1 gently stirs the cell stream, antibodies, and magnetic nanoparticles at 4°C to fully mix the cells and antibodies, so that the cells to be separated can be combined with the magnetic nanoparticles. After the combination, the cells can be attracted by the magnetic field, which facilitates the subsequent separation of the cells from the mixture.

[0031] Reference Figure 1 In the initial state, the three-way valve 14 controls the incubator 1 to be connected to the delivery pump 4. The delivery pump 4 delivers the mixture of cells and other cells that need to be separated in the incubator 1 to the multi-channel fluid separation component, and the multi-channel fluid separation component separates the cells that need to be separated from the mixture. The multi-channel fluid separation component has two outlets, one outlet is connected to the incubator 1, and the other outlet is connected to the collection chamber 7. A first control valve 12 is installed between the multi-channel fluid separation component and the incubator 1, and a second control valve 13 is installed between the multi-channel fluid separation component and the collection chamber 7. When cell separation is performed, the first control valve 12 is closed, the second control valve 13 is closed, and the cells that need to be separated are temporarily stored in the multi-channel fluid separation component. After the cells that need to be separated are separated from the mixture, the three-way valve 14 controls the delivery pump 4 to be connected to the buffer storage device 15, the first control valve 12 is opened, and the delivery pump 4 delivers the buffer to the multi-channel fluid separation component and delivers the mixture to the incubator 1. After the mixture in the multi-channel fluid separation assembly is completely drained, the first control valve 12 is closed, the second control valve 13 is opened, and the buffer solution drives the separated cells to flow into the collection chamber 7 for collection.

[0032] Reference Figure 1 and Figure 2Specifically, the multi-channel fluid separation component includes multiple channel tubes 5, coils 6 and multiple valves 8. Each channel tube 5 is connected to a temporary storage tube 11. The channel tube 5 is connected to the first control valve 12, and the temporary storage tube 11 is connected to the second control valve 13. It should be noted that the volume of the temporary storage tube 11 is similar to that of the incubator 1. The temporary storage tube 11 can accommodate the entire mixture in the incubator 1. For the convenience of display and understanding, Figure 1 In the figure, the incubator 1 is enlarged and the temporary storage tube 11 is reduced. The temporary storage tube 11 is made of an air-permeable and water-tight material such as expanded polytetrafluoroethylene.

[0033] Coil 6 is mounted on temporary storage tube 11, with each coil 6 corresponding to each temporary storage tube 11. This means that each temporary storage tube 11 is provided with a coil 6 on its outer wall. The magnetic field strength and direction of each coil 6 can be independently adjusted, allowing each channel tube 5 to be used for different cell separations. Specifically, a variable power supply can be designed for each coil 6. This variable power supply precisely controls the current flowing through the coil 6 and can be adjusted in real time, enabling dynamic adjustment of the magnetic field strength and direction based on experimental requirements.

[0034] It is understood that the coil 6 is not necessarily disposed outside the temporary storage tube 11, but may also be disposed within the wall of the temporary storage tube 11, as long as it can attract the cells in the temporary storage tube 11. The number of channel tubes 5 and coils 6 can be freely adjusted as needed and is not limited to the four shown in the figure.

[0035] In order to avoid magnetic field leakage and interference between magnetic fields generated by adjacent coils 6, a shielding cover made of magnetic shielding material (not shown in the figure) can be provided between adjacent coils 6. The magnetic shielding material can specifically be a soft magnetic alloy or a nickel-iron alloy.

[0036] Reference Figure 1 The valve 8 is set in a one-to-one correspondence with the channel pipe 5, that is, each channel pipe 5 is provided with a valve 8, and the valve 8 is set at one end of the channel pipe 5 close to the delivery pump 4 to control the conduction and closing of the channel pipe 5.

[0037] In this embodiment, the channel tube 5 is a pipe with an inner diameter of 1-3 cm and a length of 5-15 cm. The inner diameter of the coil 6 is 1.5-3.5 cm. The number of turns of the coil 6 is 20-60, and the interval between each turn is 1-3 mm.

[0038] It should be noted that the temporary storage tube 11 is arranged vertically, and the axis of the coil 6 is parallel to the axis of the temporary storage tube 11, thereby generating a magnetic field from bottom to top in the temporary storage tube 11, accurately attracting the cells to be separated in the temporary storage tube 11, and separating the cells to be separated from the mixture under the action of gravity and the magnetic field. The cells to be separated are adsorbed in the upper half of the temporary storage tube 11 due to the attraction of the magnetic field, and the mixture is in the lower half of the temporary storage tube 11 under the action of gravity, thereby achieving precise separation of the cells.

[0039] It should be noted that when the delivery pump 4 delivers the mixture to the channel tube 5, only one channel tube 5 is opened and the other channel tubes 5 are closed, that is, in the initial state, only one valve 8 is in the open state and the other valves 8 are in the closed state, so that the cells bound to the magnetic nanoparticles are separated. After completing the separation of one type of cell, the mixture is returned to the incubator 1, and magnetic nanoparticles and different antibodies are added to allow another type of cell to bind to the magnetic nanoparticles. When the delivery pump 4 delivers the mixture to the channel tube 5 again, the previously opened valve 8 is closed, and the other valve 8 is opened to separate another type of cell, and the cycle is repeated until all cells in the cell flow are separated.

[0040] It is understood that, under normal circumstances, the number of channel tubes 5 can be greater than or equal to the number of cell types that need to be separated. In this case, the equipment can be cleaned after all cells are separated, eliminating the need to repeatedly wait for cleaning of the equipment, simplifying the operation of separating multiple cells, and improving cell separation efficiency. When the number of channel tubes 5 is limited, the number of channel tubes 5 can also be less than the number of cell types that need to be separated. In this case, after separating some cells, the channel tubes 5 that have already been used for cell separation can be disassembled and cleaned while the cell separation is still in progress, which can still simplify the operation of separating cells and improve cell separation efficiency.

[0041] A counter (not shown in the figure) is provided in the collecting chamber 7 for counting the separated cells.

[0042] Reference Figure 1 The bottom end of the incubator 1 is connected to a waste liquid recovery tank 10, and the incubator 1 is connected to a cleaning pipe 9. The cleaning pipe 9 inputs cleaning liquid into the incubator 1 to clean the incubator 1. After cleaning, the waste liquid is discharged into the waste liquid recovery tank 10 for recovery. The waste liquid recovery tank 10 is also used to complete the waste liquid recovery after cell separation.

[0043] The working principle of the present invention is as follows: the cell flow and the magnetic nanoparticles with different antibodies are respectively fed into the incubator 1 through the first input tube 2 and the second input tube 3. The diameter of the magnetic nanoparticles is 50 nanometers and the concentration is 10 8Particles / mL. After the cells are combined with the magnetic nanoparticles, one of the valves 8 is opened, the first control valve 12 and the second control valve 13 are closed, and the three-way valve 14 controls the delivery pump 4 to be connected to the incubator 1, and the delivery pump 4 delivers the mixture to the temporary storage tube 11, and the magnetic field generated by the coil 6 is used to separate the cells in the temporary storage tube 11. After completing the cell separation, the first control valve 12 is opened, and the three-way valve 14 controls the buffer storage device 15 to be connected to the delivery pump 4, and the delivery pump 4 delivers buffer to the channel tube 5. Under the action of gravity, the mixture falls into the channel tube 5 and returns to the incubator 1 along with the buffer. The opened valve 8 is closed, and the other valve 8 is opened, and different antibodies are input into the incubator 1 so that different cells are combined with the magnetic nanoparticles. The above steps are repeated to separate other cells until all the cell separations are completed.

[0044] The above are all preferred embodiments of the present invention, and are not intended to limit the scope of protection of the present invention. Therefore, any equivalent changes made based on the structure, shape, and principle of the present invention should be included in the scope of protection of the present invention.

Claims

1. A multi-channel cell separation device based on magnetic nanoparticles, characterized by: The invention comprises an incubator (1), wherein the incubator (1) is connected to a first input pipe (2) and a second input pipe (3), the incubator (1) is connected to a plurality of channel pipes (5), each of the channel pipes (5) is connected to a temporary storage pipe (11), the temporary storage pipe (11) is arranged upward at one end away from the channel pipe (5), a coil (6) is provided on the temporary storage pipe (11), a first control valve (12) is provided at one end of the channel pipe (5) away from the incubator (1), a second control valve (13) is provided at one end of the temporary storage pipe (11) away from the channel pipe (5), a collecting chamber (7) is connected to the second control valve (13), and a valve (8) is provided between each of the channel pipes (5) and the incubator (1).

2. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: The first control valve (12) is connected to the incubator (1) through a pipeline.

3. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: The temporary storage tube (11) has an inner diameter of 1-3 cm and a length of 5-15 cm.

4. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: The inner diameter of the coil (6) is 1.5-3.5 cm, the number of turns of the coil (6) is 20-60 turns, and the interval between each turn is 1-3 mm.

5. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: Each of the collecting chambers (7) is provided with a counter.

6. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: A delivery pump (4) is connected between the incubator (1) and the channel tube (5).

7. The multi-channel cell separation device based on magnetic nanoparticles according to claim 6, characterized in that: A three-way valve (14) is connected between the delivery pump (4) and the incubator (1), and a buffer storage device (15) is connected to the three-way valve (14).

8. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: The incubator (1) is connected to a cleaning pipe (9).

9. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: The incubator (1) is connected to a waste liquid recovery tank (10).

10. The multi-channel cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: The temporary storage tube (11) is arranged vertically.

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