Cell separation device and analysis device based on magnetic nanoparticles

Abstract

The utility model provides a cell separation device and an analysis device based on magnetic nanoparticles, and belongs to the technical field of biomedicine. The separation device comprises a cell and magnetic nanoparticle incubation unit, a cell separation unit and a cell collection unit. Target cells are accurately captured mainly through magnetic nanoparticles with antibodies and are accurately separated out through magnetic field adsorption force of an electromagnetic coil. According to the separation device and the analysis device, the whole multi-channel design in the cell incubation, separation, collection and analysis processes simultaneously realizes the treatment of a plurality of samples and the synchronous separation of different cells, and the accuracy of cell separation is improved through accurate magnetic field control and magnetic nanoparticles. The requirements on high throughput and high precision of cell separation at present are met.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the field of biomedical technology, more particularly to a separation device and analysis device based on magnetic nanoparticles. BACKGROUND

[0002] Cell separation technology plays an important role in biomedical research and clinical applications. Cell separation technology refers to the technology of separating target cells from other cells or impurities based on the physical, chemical or biological characteristics of cells from biological samples (such as tissues, cell culture medium, etc.). Common cell separation methods include flow cytometry sorting, immunomagnetic bead separation and centrifugal separation. These methods have played an important role in the capture and collection of specific cells, but also have some significant limitations.

[0003] Flow cytometry sorting: can perform high-precision analysis and sorting on single cells, but its equipment is expensive and has high maintenance cost, the operation is complex, and professional technicians are needed to operate, and it can only handle one sample at a time, and cannot efficiently handle a large number of samples or simultaneously separate multiple cell types.

[0004] Immunomagnetic bead separation: uses functionalized magnetic beads for high-specificity separation of specific cells. Although this method has advantages in specificity and diversity of cell separation, it is usually designed as a single channel, with complex operation steps, high cost and slow processing speed, which cannot meet the high-throughput demand.

[0005] Centrifugal separation: uses the difference in cell size and density to separate cells by gradually changing the centrifugal speed. The device is simple and suitable for separating large volume samples, but the separation precision is not high, and cell damage is easy to occur, especially under high speed conditions. It is not suitable for fine cell type separation or simultaneous separation of multiple cell types.

[0006] In summary, the above-mentioned cell separation technologies at least have the following technical problems:

[0007] 1. Limited throughput and low efficiency: Most existing devices can only handle a single sample or a single cell type, making it difficult to meet the demand for simultaneous separation of multiple cell types in modern high-throughput research or clinical applications.

[0008] 2. Complex operation and high cost: For example, flow cytometry sorting and immunomagnetic bead separation require expensive equipment and consumables, and the operation steps are complex, increasing the time and cost of experiments.

[0009] 3. Cell damage and separation precision problems: For example, centrifugal separation is prone to mechanical damage to cells and cannot achieve high-precision cell type separation, which is particularly disadvantageous for research that requires cell activity and function to be maintained. UTILITY MODEL CONTENT

[0010] In order to solve the problems in the prior art and meet the requirements of high throughput and high precision for cell separation, the utility model provides a separation device and analysis device based on magnetic nanoparticles.

[0011] In order to achieve the above technical purpose, the utility model adopts the following technical scheme:

[0012] The utility model provides a separation device based on magnetic nanoparticles, including cell and magnetic nanoparticle incubation unit, cell separation unit, cell collection unit,

[0013] The cell and magnetic nanoparticle incubation unit includes a plurality of cell incubators with consistent arrangement directions, each cell incubator includes a cell storage container, a rotating mechanism and a fixing mechanism, the rotating mechanism is fixedly connected to the fixing mechanism, the cell storage container is rotatably connected to the fixing mechanism, the output end of the rotating mechanism is connected to the cell storage container, the cell storage container can rotate with the rotating mechanism, the cell storage container is provided with a cell incubation inlet and a cell incubation outlet, and each cell incubation outlet is provided with a separate micro pump.

[0014] The cell separation unit includes a plurality of micro flow channels, each micro flow channel is provided with a cell separation inlet and a cell separation outlet, a plurality of cell separation inlets are connected to a plurality of cell incubation outlets in one-to-one correspondence, and a separate electromagnetic coil is arranged outside each micro flow channel;

[0015] The cell collection unit includes a plurality of cell collection devices, each cell collection device is provided with a cell collection inlet and a cell collection outlet, a plurality of cell collection inlets are connected to a plurality of cell separation outlets in one-to-one correspondence through a three-way valve, and the other outlet of the three-way valve is connected to a waste liquid recovery device through a blowdown pipeline.

[0016] Preferably, the electromagnetic coils are connected to independent power supplies.

[0017] Preferably, each electromagnetic coil can independently set the magnetic field strength and direction.

[0018] Preferably, the cell collection unit is connected to a storage temperature control system.

[0019] The utility model also provides an analysis device based on magnetic nanoparticles: including the above-mentioned cell separation device and cell analysis unit, the input of the cell analysis unit is connected with the cell collection unit of the cell separation device.

[0020] Preferably, the cell analysis unit includes a resistance pulse counter and a high-speed camera, and a plurality of cell collection outlets are connected to the resistance pulse counter.

[0021] Compared with the prior art, the utility model has the beneficial effects that:

[0022] 1. The multi-channel design of the whole process from cell incubation, separation, collection to analysis can simultaneously process multiple samples and synchronously separate multiple cells, meeting the needs of cell separation diversity, greatly improving the throughput and overall efficiency of cell separation. The multi-channel can be used only partially or increased according to actual needs, and the flexibility and utilization rate of the device are high. Moreover, the overall structure of the device is simple, and the maintenance cost is low.

[0023] 2. The accuracy of cell separation is improved by precise magnetic field control and magnetic nanoparticles, and mechanical damage to cells is avoided, maintaining the activity and function of the cells. At the same time, the rotatable design of the cell incubator can enhance the uniformity of cell and nanoparticle incubation.

[0024] 3. By increasing the cell analysis unit, the separated cells can be analyzed in real time, and the number of cells can be counted, enhancing the practicality and data acquisition ability of the device, and simplifying the subsequent operation steps and analysis cost. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 It is a structural schematic view of an embodiment of the cell separation device and analysis device of the present application.

[0026] Figure 2 It is a structural schematic view of the cell incubator.

[0027] In the drawings: 1, three-way valve; 2, micro pump; 3, electromagnetic coil; 4, cell collection device; 5, waste liquid recovery device; 6, cell incubator; 6.1, cell storage container; 6.11, cell incubation inlet; 6.12, cell incubation outlet; 6.13, magnetic nanoparticles; 6.2, fixing mechanism; 6.3, rotating mechanism; 6.31, gear disc; 7, drain line, 8, buffer solution line. DETAILED DESCRIPTION

[0028] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of the present application.

[0029] In the description of the utility model, it is understood that the orientation or positional relationship indicated by the terms "coaxial", "bottom", "one end", "top", "middle", "the other end", "upper", "one side", "top", "inner", "front", "central", "both ends" and the like is based on the orientation or positional relationship shown in the drawings, and is only for the convenience of describing the utility model and simplifying the description, and therefore cannot be understood as a limitation on the utility model.

[0030] In the utility model, unless otherwise expressly specified and limited, the terms "mounting", "setting", "connection", "fixing", "screw connection" and the like should be understood in a broad sense, for example, can be fixed connection, or can be detachable connection, or can be integrated; can be mechanical connection, or can be electrical connection; can be directly connected, or can be indirectly connected through an intermediate medium; can be the communication inside two elements or the interaction relationship of two elements, unless otherwise expressly limited, the above-mentioned terms in the utility model can be understood according to the specific meaning in the utility model by the person skilled in the art according to the specific situation.

[0031] As shown in the accompanying Figure 1 The utility model discloses an analysis device and separation device based on magnetic nano particle, and provides a kind of based on magnetic nano particle separation device, including: including cell and magnetic nano particle incubation unit, cell separation unit, cell collection unit;

[0032] Cell and magnetic nano particle incubation unit includes a plurality of cell incubators 6 of consistent arrangement direction, each cell incubator includes cell storage container 6.1, rotating mechanism 6.3 and fixed mechanism 6.2, rotating mechanism 6.3 is fixedly connected to fixed mechanism 6.2, cell storage container 6.1 is rotatably connected to fixed mechanism 6.2, and cell storage container 6.1 is provided with gear disc 6.31;Rotating mechanism 6.3 output end is connected with cell storage container 6.1 by meshing with gear disc 6.31 and is drivingly connected;Cell storage container 6.1 can rotate with rotating mechanism 6.3, and cell storage container 6.1 is provided with cell incubation inlet 6.11 and cell incubation outlet 6.12, and each cell incubation outlet 6.12 is provided with a separate micro pump 2;Cell storage container 6.1 is loaded with magnetic nano particle 6.13.

[0033] Cell separation unit includes a plurality of microchannels, each microchannel is provided with cell separation inlet and cell separation outlet, a plurality of cell separation inlets are connected with a plurality of cell incubation outlets 6.12 one by one, and a separate electromagnetic coil 3 is provided outside each microchannel;

[0034] The cell collection unit comprises a plurality of cell collection devices 4, each of which is provided with a cell collection inlet and a cell collection outlet, the plurality of cell collection inlets are connected with the plurality of cell separation outlets one by one through a three-way valve 1, and the other outlet of the three-way valve is connected with the waste liquid recovery device 5 through a waste liquid pipeline 7. Here, one end of the outlet of the three-way valve 1 is connected with the cell collection device, and the other end of the outlet is connected with the waste liquid recovery device, and the purpose of collecting the purified target cells by the cell collection device can be realized by controlling the outlet of the valve, and the non-target cells are recovered into the waste liquid recovery device as waste liquid in the purification process.

[0035] In the embodiment, the cell and magnetic nanoparticle incubation unit is provided with a plurality of parallel arranged cell incubators, each of which has a capacity of 10 milliliters, and the diameter of the magnetic nanoparticle 6.13 is between 10 nanometers and 100 nanometers. In the cell separation unit, the inner diameter of the microchannel is 1-3 centimeters, and the length is 5-15 centimeters; the inner diameter of the coil is 1.5-3.5 centimeters, 20-100 turns, and the interval between each turn of the adjacent coil is 0-3 millimeters.

[0036] In some embodiments of the utility model, each electromagnetic coil is connected with independent power supply, and each electromagnetic coil can individually adjust its magnetic field intensity and direction. Specifically, the magnetic field intensity is adjusted by adjusting the current size and the number of turns of the coil, and the magnetic field direction is adjusted by adjusting the current direction and the winding direction of the coil.

[0037] In some embodiments of the utility model, the cell collection unit is connected with a storage temperature control system. The control temperature range is 4 DEG C to 37 DEG C, so as to keep the cell viability.

[0038] The utility model also provides a kind of separation device based on magnetic nanoparticle: including the cell separation device implementation way and cell analysis unit of any one described above, the input of cell analysis unit is connected with the cell collection unit of cell separation device.

[0039] Preferably, the cell analysis unit includes a resistance pulse counter and a high-speed camera, and the plurality of cell collection outlets are connected with the resistance pulse counter, which can count and analyze the collected cell information in real time through the resistance pulse counter and the high-speed camera, and generate a data file for transmission to an external computer.

[0040] In some embodiments of the utility model, the cell analysis unit can be configured with a high-speed camera to record the morphology of the collected cells. The high-speed camera has a resolution of 2048x2048 pixels and a frame rate of up to 1000 frames per second. In some embodiments of the utility model, the separation device and the analysis device can use only part of the channels as needed, or increase the channels as needed to separate more sample cells at the same time.

[0041] The whole process of each channel cell incubation, separation, collection and analysis of the separation device and analysis device is as follows:

[0042] 1. Cell incubation process: in the cell and magnetic nanoparticle incubation unit, the surface of the magnetic nanoparticle 6.13 is functionalized to have specific antibodies corresponding to target cells (such as specific immune cells or tumor cells and various types of cells). The sample cell liquid enters the corresponding cell incubator for incubation. The rotation of the cell incubator makes the incubation more uniform, and the magnetic nanoparticles 6.13 with corresponding antibodies are adsorbed on the target cells through immune reaction, so that the target cells are attached with magnetism. The magnetic nanoparticles 6.13 can be designed as nanoparticles with different antibody types, which can separate different target cells in different pipelines in an external magnetic field.

[0043] 2. Cell separation and collection process: first, close the outlet valve of the three-way valve to form a closed space in the cell separation unit, and the electromagnetic coil is connected to the power supply. After the pump is started, a pressure difference is generated at the outlet of the cell incubator, which promotes the directional flow of the sample cell liquid to the cell separation unit. Under the action of the magnetic field of the electromagnetic coil, the target cells with magnetism are adsorbed to the wall. Then open the outlet valve leading to the waste liquid recovery device, so that the non-target cells without antibody binding flow into the waste liquid recovery device through the drain pipe. After the waste liquid is discharged, turn the three-way valve to the cell collection device, and the electromagnetic coil is de-energized to make the magnetic field disappear. Then, the buffer solution is injected through the buffer solution pipeline 8 to flow the target cells that need to be recovered and previously adsorbed on the wall into the cell collection device for storage.

[0044] The target cells carrying magnetism are adsorbed and separated accurately. The purified target cells enter the collection unit for collection and storage.

[0045] 3. Cell analysis process: the purified target cells collected can subsequently enter the cell analysis unit when analysis is required. The collected cell information can be counted and analyzed in real time by resistance pulse counter and high-speed camera.

[0046] The multi-channel design can separate different specific cells of different samples at the same time, and each channel is independent of each other. Different magnetic nanoparticles 6.13 with corresponding antibodies can be incubated in the cell and magnetic nanoparticle incubation unit of the corresponding channel; for different target cells, the magnetic field strength and direction of the corresponding channel are adjusted to improve the accuracy of separation. The cell separation and analysis device realizes high-precision and high-throughput separation of different cells at the same time.

[0047] The above merely describes the embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement and improvement within the scope of the present application should be included in the protection scope of the present application.

Claims

1. A cell separation device based on magnetic nanoparticles, characterized by: The device comprises a cell and magnetic nanoparticle incubation unit, a cell separation unit, and a cell collection unit. The cell and magnetic nanoparticle incubation unit comprises a plurality of cell incubators arranged in the same direction, each of which comprises a cell storage container, a rotating mechanism, and a fixing mechanism. The rotating mechanism is fixedly connected to the fixing mechanism, and the cell storage container is rotatably connected to the fixing mechanism. The output end of the rotating mechanism is connected to the cell storage container.

2. The cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: The cell storage container is rotatable with the rotating mechanism.

3. The cell separation device based on magnetic nanoparticles according to claim 2, characterized in that: The cell storage container is provided with a cell incubation inlet and a cell incubation outlet.

4. The cell separation device based on magnetic nanoparticles according to claim 1, characterized in that: Each cell incubation outlet is provided with a separate micro pump.

5. A cell analysis device based on magnetic nanoparticles, characterized by: The cell separation unit comprises a plurality of micro channels.

6. The magnetic nanoparticle-based cellular analysis device of claim 5, wherein: Each micro channel is provided with a cell separation inlet and a cell separation outlet. A plurality of cell separation inlets are connected to a plurality of cell incubation outlets one by one. An individual electromagnetic coil is provided outside each micro channel. The electromagnetic coils are connected to independent power sources. Each electromagnetic coil can independently set the magnetic field strength and direction through an independent power source. The cell collection unit is connected to a storage temperature control system. The device comprises a cell separation device according to any one of claims 1-4 and a cell analysis unit. The input of the cell analysis unit is connected to the cell collection unit of the cell separation device. The cell analysis unit comprises a resistance pulse counter and a high-speed camera. A plurality of cell collection outlets are connected to the resistance pulse counter.

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