CN115127977A - Magnetic beads and methods of making the same - Google Patents

Abstract

The present application relates to the technical field of magnetic bead modification, and specifically discloses a magnetic bead and a manufacturing method thereof. The magnetic bead comprises: a magnetic microsphere, including a core microsphere, and a first magnetic nanoparticle coupled to the outer surface of the core microsphere and a first coating layer coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle; the second magnetic nanoparticle is coupled to the outer surface of the first coating layer; the third magnetic nanoparticle is at least dispersed Distributed inside the first cladding layer and/or inside the gel layer. Through the above method, while improving the magnetic response of the magnetic beads, the interference signal caused by impurities can be reduced, the autofluorescence of the microspheres can be reduced, and the detection sensitivity can be improved.

Description

Magnetic beads and methods of making the same

technical field

The present application relates to the technical field of magnetic bead modification, and in particular, to a magnetic bead and a manufacturing method thereof.

Background technique

Magnetic microspheres are a kind of spherical composite materials with a diameter of nanometer or micrometer. The magnetic microspheres are composed of microspheres and magnetic nanoparticles adsorbed on the surface of the microspheres.

In the long-term research and development process, the inventors of the present application found that if the surface of the microspheres adsorbs magnetic nanoparticles excessively, as shown in Figure 26, the impurities on the magnetic microspheres will increase, and the signal of the impurities will seriously interfere with the main flow cytometer on the flow cytometer. In addition, heating is required in the process of coating the polymer protective layer, which easily causes the microspheres to generate strong autofluorescence, resulting in a decrease in detection sensitivity.

SUMMARY OF THE INVENTION

The present application aims to solve at least one of the technical problems existing in the prior art. Therefore, one purpose of the present application is to propose a magnetic bead and a manufacturing method thereof, which can improve the magnetic response of the magnetic bead, reduce the interference signal caused by impurities, reduce the autofluorescence of the microsphere, and improve the detection sensitivity.

A first aspect of the present application provides a magnetic bead. The magnetic bead includes: a magnetic microsphere, including a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a first magnetic nanoparticle coated on the outer surface of the core microsphere and a first magnetic nanoparticle. A first coating layer on the outer surface of the magnetic nanoparticle; a second magnetic nanoparticle coupled to the outer surface of the first coating layer; and a third magnetic nanoparticle dispersed and distributed at least inside the first coating layer.

A second aspect of the present application provides a method for manufacturing a magnetic bead, the method comprising: providing a magnetic microsphere, the magnetic microsphere comprising a core microsphere, first magnetic nanoparticles coupled to the outer surface of the core microsphere, and a package A first coating layer covering the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle; the magnetic microsphere and the second magnetic nanoparticle are combined by adsorption technology, so that the second magnetic nanoparticle is coupled to the first coating The outer surface of the layer; wherein, before the step of combining magnetic microspheres and second magnetic nanoparticles using adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer, or before adopting adsorption technology After the step of combining the magnetic microspheres and the second magnetic nanoparticles so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer, the method further includes: combining the magnetic microspheres and the third magnetic nanoparticles by a swelling technique nanoparticles, so that the third magnetic nanoparticles are dispersed and distributed in the interior of the first coating layer and/or the interior of the gel layer.

Different from the situation in the prior art, the magnetic beads of the present application include: magnetic microspheres, second magnetic nanoparticles and third magnetic nanoparticles, and the magnetic beads include: magnetic microspheres, including core microspheres, coupled to the core the first magnetic nanoparticle on the outer surface of the microsphere and the first coating layer coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle; the second magnetic nanoparticle is coupled to the outer surface of the first coating layer ; The third magnetic nanoparticles are at least dispersed in the interior of the first coating layer, as shown in Figure 27, under the premise of avoiding excessive adsorption of magnetic nanoparticles on the surface of the magnetic microspheres, the first coating layer swells through the interior of the first coating layer. The three magnetic nanoparticles and the externally coupled second magnetic nanoparticles can enhance the magnetic response signal of the magnetic beads, reduce the interference signal caused by impurities, reduce the autofluorescence of the magnetic microspheres, and improve the detection sensitivity.

Additional aspects and advantages of the present application will be set forth, in part, from the following description, and in part will become apparent from the following description, or may be learned by practice of the present application.

Description of drawings

The above and/or additional aspects and advantages of the present application will become apparent and readily understood from the following description of embodiments in conjunction with the accompanying drawings, wherein:

1 is a schematic flowchart of a method for making magnetic beads according to the first embodiment of the present application;

2 is a schematic flowchart of a method for making magnetic beads according to a second embodiment of the present application;

3 is a schematic flowchart of a method for making magnetic beads according to a third embodiment of the present application;

Fig. 4 is a schematic flowchart of step S10 in Figs. 1-3;

5 is a schematic flowchart of a method for manufacturing a magnetic bead according to a fourth embodiment of the present application;

6 is a schematic flowchart of a method for making magnetic beads according to the fifth embodiment of the present application;

FIG. 7 is a schematic flowchart of a method for making magnetic beads according to the sixth embodiment of the present application;

8 is a schematic flowchart of a method for making magnetic beads according to the seventh embodiment of the present application;

9 is a schematic flowchart of a method for manufacturing a magnetic bead according to an eighth embodiment of the present application;

10 is a schematic flowchart of a method for making magnetic beads according to the ninth embodiment of the present application;

11 is a schematic flowchart of a method for producing magnetic beads according to the tenth embodiment of the present application;

12 is a schematic flowchart of a method for manufacturing a magnetic bead according to the eleventh embodiment of the present application;

13 is a schematic flowchart of a method for making magnetic beads according to the twelfth embodiment of the present application;

14 is a schematic flowchart of a method for producing magnetic beads according to the thirteenth embodiment of the present application;

15 is a schematic flowchart of a method for manufacturing a magnetic bead according to the fourteenth embodiment of the present application;

16 is a schematic flowchart of a method for making magnetic beads according to the fifteenth embodiment of the present application;

17 is a schematic flowchart of a method for making magnetic beads according to the sixteenth embodiment of the present application;

18 is a schematic flowchart of a method for manufacturing a magnetic bead according to the seventeenth embodiment of the present application;

19 is a schematic flowchart of a method for making magnetic beads according to the eighteenth embodiment of the present application;

Fig. 20 is a schematic flowchart of step S30 in Fig. 1-Fig. 19;

21 is a schematic diagram of the first structure of the magnetic bead proposed in the present application;

Fig. 22 is the second structural schematic diagram of the magnetic bead proposed by the present application;

23 is a schematic diagram of a method for making magnetic microspheres proposed by the present application;

Fig. 24 is the third structural schematic diagram of the magnetic bead proposed by the present application;

FIG. 25 is a schematic diagram of the fourth structure of the magnetic bead proposed by the present application;

Fig. 26 is the magnetic response signal diagram when the magnetic nanoparticles are excessively adsorbed on the surface of the microsphere in the prior art;

Fig. 27 is the magnetic response signal diagram of the magnetic beads of the present application;

Fig. 28 is the fifth structural schematic diagram of the magnetic bead proposed by the present application;

FIG. 29 is a schematic diagram of the sixth structure of the magnetic bead proposed in the present application.

Detailed ways

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present application.

Throughout the specification, unless specifically stated otherwise, terms used herein are to be understood as commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. In case of conflict, the present specification takes precedence.

It should be noted that, in the embodiments of the present application, the terms "comprising", "comprising" or any other variations thereof are intended to cover non-exclusive inclusion, so that a method or device including a series of elements not only includes the explicitly stated elements, but also other elements not expressly listed or inherent to the implementation of the method or apparatus. Without further limitation, an element defined by the phrase "comprising a..." does not preclude the presence of additional related elements (eg, steps in a method or elements in an apparatus) in the method or apparatus that includes the element , where a unit may be part of a circuit, part of a processor, part of a program or software, etc.).

It should be noted that the term "first\second\third" involved in the embodiments of the present application is only to distinguish similar objects, and does not represent a specific ordering of objects. It is understandable that "first\second\third" "Three" may be interchanged in a particular order or sequence where permitted. It should be understood that the "first\second\third" distinctions may be interchanged under appropriate circumstances, so that the embodiments of the present application described herein can be implemented in sequences other than those illustrated or described herein.

Referring to FIG. 1 , the first embodiment of the present application proposes a method for making magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

The above-mentioned first magnetic nanoparticles may be water-soluble magnetic nanoparticles.

Optionally, the outer surface of the core microsphere can be modified by chemical means to the required first charged functional group, wherein the first charged functional group includes at least one of a charged carboxyl group, an amino group, a sulfonic acid group or a sulfhydryl group, The possibility of coupling the first magnetic nanoparticles to the core microspheres is provided.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

Specifically, the second magnetic nanoparticles can be water-soluble magnetic nanoparticles, and the coupling between the first coating layer and the second magnetic nanoparticles can be achieved by adsorption, van der Waals force and/or covalent bonding.

Optionally, the outer surface of the first coating layer can be modified by chemical means to the required second charged functional group, wherein the second charged functional group includes at least one of a charged carboxyl group, an amino group, a sulfonic acid group or a sulfhydryl group. This provides the possibility for the first coating layer to couple the second magnetic nanoparticles.

Dissolve the second magnetic nanoparticles in deionized water, add magnetic microspheres, and cross-link the second magnetic nanoparticles with the magnetic microspheres, so that the outer surface of the core microspheres is coupled with the second magnetic nanoparticles .

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Mixing the magnetic microspheres prepared in step S20 with the second magnetic nanoparticles coupled on the outer surface and the gel material, adding a cross-linking agent, stirring the reaction, and after the obtained product is allowed to stand, magnetic separation is performed to remove the residual gel material, Microspheres whose outer surface is covered with a gel layer are obtained. Wherein, the gel layer covers at least part of the second magnetic nanoparticles.

Wherein, the material of the gel layer can be at least one of chitosan, sodium alginate, polyacrylic acid, polymethacrylic acid, polyacrylamide, and polyN-polyacrylamide.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by using the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed in the inside of the first coating layer and/or the inside of the gel layer.

Specifically, the third magnetic nanoparticles may be fat-soluble magnetic nanoparticles.

Disperse the magnetic microspheres in the first medium, provide a second medium containing the third magnetic nanoparticles, mix, and then perform a swelling reaction (the vortex is dispersed uniformly, and the rotation reaction time is preset), and the third magnetic nanoparticles are packaged. The magnetic microspheres are embedded in the interior of the magnetic microspheres and/or the gel layer, and the supernatant liquid is magnetically separated to obtain the microspheres with the third magnetic nanoparticles distributed inside. Wherein, the first medium and the second medium are both swelling media, specifically a single or mixed solvent that can not only disperse and distribute the third magnetic nanoparticles, but also swell magnetic microspheres, for example, the swelling medium can be chloroform, two One or a combination of methyl chloride, ethanol, methanol, hexanediol, n-butanol, isobutanol, n-hexane, cyclohexane, and tetrahydrofuran, but not limited to the above-mentioned substances. It can be understood that, after the third magnetic nanoparticles enter the interior of the core microspheres through swelling, the third magnetic nanoparticles are embedded in the interior of the core microspheres. Further, during the process of embedding the lipid-soluble magnetic nanoparticles and the lipid-soluble fluorescent dyes into the core microspheres, the lipid-soluble magnetic nanoparticles and the lipid-soluble fluorescent dyes can be embedded in the interior of the gel layer.

Referring to FIG. 2 , a second embodiment of the present application proposes a method for making magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

The above-mentioned first magnetic nanoparticles may be water-soluble magnetic nanoparticles.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

The above-mentioned third magnetic nanoparticles may be fat-soluble magnetic nanoparticles.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

The above-mentioned second magnetic nanoparticles are water-soluble magnetic nanoparticles.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Different from the first embodiment, the second embodiment “combines magnetic microspheres and third magnetic nanoparticles using swelling technology” occurs before “combining magnetic microspheres and second magnetic nanoparticles using adsorption technology”, so , which can avoid the difficulty of the fat-soluble magnetic nanoparticles entering the interior of the magnetic microspheres due to the hydrophilicity (ie oleophobicity) of the water-soluble magnetic nanoparticles, and greatly improve the encapsulation of the lipid-soluble magnetic nanoparticles in the magnetic microspheres. Buried quantity.

Referring to FIG. 3 , a third embodiment of the present application proposes a method for making magnetic beads. The method includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

The above-mentioned first magnetic nanoparticles may be water-soluble magnetic nanoparticles.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

The above-mentioned second magnetic nanoparticles may be water-soluble magnetic nanoparticles.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

The above-mentioned third magnetic nanoparticles may be fat-soluble magnetic nanoparticles.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Different from the first embodiment, the “combination of magnetic microspheres and third magnetic nanoparticles using swelling technology” in the third embodiment occurs before the coating of the gel layer, so the third magnetic nanoparticles are more likely to enter the core microparticles. Inside the sphere, the embedding quantity of the third magnetic nanoparticles in the magnetic microsphere is greatly increased.

Optionally, the gel layer in step S30 of the first embodiment to the third embodiment can be replaced with a second polymer coating layer, and the material of the second polymer coating layer is polystyrene, polymethacrylic acid At least one of methyl ester, polyethylene-methyl methacrylate, polyacrylonitrile, polyethylene, polypropylene, and polyethyl acrylate.

Thus, the structure of the magnetic bead 10 prepared by the above-mentioned first to third embodiments or their alternatives is shown in FIG. 21 or 22 , and the magnetic bead 10 includes: magnetic microspheres 11 and second magnetic nanoparticles 12 , the third magnetic nanoparticles 13 and the second coating layer 14 . The magnetic microsphere 11 includes a core microsphere 111 , a first magnetic nanoparticle 112 coupled to the outer surface of the core microsphere 111 , and a first magnetic nanoparticle 112 coated on the outer surface of the core microsphere 111 and the outer surface of the first magnetic nanoparticle 112 . A cladding layer 113 . The second magnetic nanoparticles 12 may be coupled to the outer surface of the core microspheres 111 . The second coating layer 14 coats the outer surfaces of the core microspheres 111 and the outer surfaces of the first magnetic nanoparticles 112 . Optionally, the second coating layer 14 coats all the water-soluble magnetic nanoparticles, and the outer surface of the magnetic beads 10 is a smooth surface. The third magnetic nanoparticles 13 are dispersed and distributed at least inside the first coating layer 113. It can be understood that, after the third magnetic nanoparticles 13 enter the interior of the core microspheres 111 through swelling, the third magnetic nanoparticles 13 are embedded in the core microspheres 111. The interior of the core microsphere 111 . Wherein, the above-mentioned coating layer 14 may be a gel layer or a second polymer coating layer. It can be understood that the third magnetic nanoparticles 13 can be dispersed in the gel layer, and limited by the material of the second polymer coating layer, the third magnetic nanoparticles 13 cannot be dispersed in the second polymer coating. The interior of the cladding.

Further, step S30 in the above-mentioned first embodiment to third embodiment can be deleted, and the structure of the magnetic bead 10 thus obtained is shown in FIG. 28 . The magnetic bead 10 includes: a magnetic microsphere 11 , a second Magnetic nanoparticles 12 and third magnetic nanoparticles 13 . The magnetic microsphere 11 includes a core microsphere 111 , a first magnetic nanoparticle 112 coupled to the outer surface of the core microsphere 111 , and a first magnetic nanoparticle 112 coated on the outer surface of the core microsphere 111 and the outer surface of the first magnetic nanoparticle 112 . A cladding layer 113 . The second magnetic nanoparticles 12 may be coupled to the outer surface of the core microspheres 111 .

Different from the situation in the prior art, the magnetic beads of the present application include: magnetic microspheres, second magnetic nanoparticles and third magnetic nanoparticles, and the magnetic beads include: magnetic microspheres, including core microspheres, coupled to the core the first magnetic nanoparticle on the outer surface of the microsphere and the first coating layer coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle; the second magnetic nanoparticle is coupled to the outer surface of the first coating layer ; The third magnetic nanoparticles are at least dispersed in the interior of the first coating layer, as shown in Figure 27, under the premise of avoiding excessive adsorption of magnetic nanoparticles on the surface of the magnetic microspheres, the first coating layer swells through the interior of the first coating layer. The three magnetic nanoparticles and the externally coupled second magnetic nanoparticles can enhance the magnetic response signal of the magnetic beads, reduce the interference signal caused by impurities, reduce the autofluorescence of the magnetic microspheres, and improve the detection sensitivity.

Optionally, the core microspheres 111 are silica core microspheres 111 .

Optionally, the first coating layer 113 is a first polymer coating layer 113 .

Specifically, the material of the first polymer coating layer 113 is an organic monomer molecular polymer, which may include polystyrene, polymethyl methacrylate, polyethylene-methyl methacrylate, polyacrylonitrile, polyacrylonitrile At least one of ethylene, polypropylene, and polyethyl acrylate.

In some embodiments, the outer surface of the first coating layer 113 contains a second charged functional group, and the second charged functional group includes at least one of a charged carboxyl group, an amino group, a sulfonic acid group or a thiol group. The first coating layer 113 couples the second magnetic nanoparticles 12 through charged functional groups.

In some embodiments, the above-mentioned water-soluble magnetic nanoparticles are paramagnetic nanoparticles, wherein the paramagnetic nanoparticles can be selected from ferric tetroxide, ferric oxide, and alloy-type paramagnetism containing nickel or cobalt at least one of the magnetic particles.

In some embodiments, the above-mentioned fat-soluble magnetic nanoparticles are nanoparticles with paramagnetic properties, wherein the nanoparticles with paramagnetic properties can be selected from at least one of ferric oxide or ferric oxide. In addition, the fat-soluble magnetic nanoparticles contain a fat-soluble ligand of at least one of unsaturated fatty acids, saturated fatty acids, unsaturated fatty amines or saturated fatty amines. The above-mentioned lipid-soluble ligands can bind to the surface of the lipid-soluble magnetic nanoparticles, thereby stabilizing the lipid-soluble magnetic nanoparticles. Wherein, the fat-soluble ligand may include at least one of oleic acid, oleylamine or stearic acid.

In some embodiments, the particle size of the magnetic microspheres 11 is 1 μm˜50 μm (eg 1 μm, 5 μm, 10 μm, 20 μm, 50 μm), and the particle size of the third magnetic nanoparticles 13 is 1 nm˜200 nm (eg 1 nm, 5 nm, 50 nm, 100 nm, 200 nm), the particle size of the second magnetic nanoparticles 12 is 1 nm˜200 nm (for example, 1 nm, 5 nm, 50 nm, 100 nm, 200 nm).

In some embodiments, by mass percentage, the magnetic beads 10 include: 50%-99.5% of the magnetic microspheres 11, 0.1%-49.9% of the second magnetic nanoparticles 12, 0.1%-49.9% of the third magnetic Nanoparticles 13. The applicant found that within the above numerical range, the magnetic response of the magnetic beads can be improved, the interference signal caused by impurities can be reduced, and the magnetic microspheres can be reduced under the premise of avoiding excessive adsorption of magnetic nanoparticles on the surface of the magnetic microspheres. Autofluorescence improves detection sensitivity.

Referring to FIG. 4, in some embodiments, the above step S10 includes the following steps:

S11: Combining the core microspheres and the first magnetic nanoparticles using adsorption technology, so that the first magnetic nanoparticles are coupled to the outer surface of the core microspheres.

Specifically, the core microspheres are silica core microspheres. Further, the silica core microspheres can be hollow mesoporous silica core microspheres. The charged functional group required for modification, wherein the charged functional group includes at least one of charged carboxyl group, amino group, sulfonic acid group or sulfhydryl group, provides the possibility for the core microsphere to couple the first magnetic nanoparticles.

In other embodiments, a soluble ferric ion salt can be used as a raw material, dissolved in an ethylene glycol solution, and subjected to a solvothermal reaction to prepare ferric oxide magnetic nanoparticles, and then the prepared ferric oxide magnetic nanoparticles particles, using tetraethyl orthosilicate in the participation of ammonia water to synthesize silica core microspheres coupled with ferric oxide magnetic nanoparticles.

S12: Coating a first coating layer on the outer surface of the core microsphere coupled with the first magnetic nanoparticles, and the first coating layer coating at least part of the first magnetic nanoparticles.

Specifically, the first coating layer is a polymer first coating layer, and the material of the polymer first coating layer is polystyrene, polymethyl methacrylate, polyethylene-methyl methacrylate, polypropylene At least one of nitrile, polyethylene, polypropylene, and polyethyl acrylate. Optionally, the outer surface of the first coating layer can be modified by chemical means with required charged functional groups, wherein the charged functional groups include at least one of a charged carboxyl group, an amino group, a sulfonic acid group or a sulfhydryl group, which is the first A coating layer offers the possibility to couple a second magnetic nanoparticle.

Referring to FIG. 5 , a fourth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

Specifically, the magnetic microspheres are dispersed in the first medium, a third medium containing a fat-soluble fluorescent dye is provided, mixed, and then a swelling reaction is performed (the vortex is dispersed uniformly, and the rotation reaction time is preset), and the fat-soluble fluorescent dye is The dye is embedded in the core microspheres, and the supernatant is magnetically separated to obtain fluorescent microspheres with lipid-soluble fluorescent dyes distributed inside. Wherein, the first medium and the third medium are both swelling media, specifically a single or mixed solvent that can not only disperse the lipid-soluble magnetic nanoparticles, but also swell the magnetic microspheres, for example, the swelling media can be chloroform, two One or a combination of methyl chloride, ethanol, methanol, hexanediol, n-butanol, isobutanol, n-hexane, cyclohexane, and tetrahydrofuran, but not limited to the above-mentioned substances.

Further, step S50 may be performed multiple times, respectively combining lipid-soluble fluorescent dyes with different fluorescence characteristics and magnetic microspheres, or combining lipid-soluble fluorescent dyes and magnetic microspheres with different concentrations, respectively, to obtain a plurality of different fluorescent dyes. Intensity of microspheres, namely fluorescently encoded microspheres. Wherein, different lipid-soluble fluorescent dyes can be mixed in different proportions to prepare different encoded microspheres. It is precisely due to the different ratios of different lipid-soluble fluorescent dyes that endow the fluorescently encoded microspheres with different fluorescent characteristics.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by using the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed in the inside of the first coating layer and/or the inside of the gel layer.

Referring to FIG. 6 , a fifth embodiment of the present application proposes a method for making magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by using the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed in the inside of the first coating layer and/or the inside of the gel layer.

Referring to FIG. 7 , a sixth embodiment of the present application proposes a method for making magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

S50: Using swelling technology to combine magnetic microspheres and liposoluble fluorescent dyes, the liposoluble fluorescent dyes are dispersed and distributed in the interior of the first coating layer and/or the interior of the gel layer.

Further, in the process of embedding the lipid-soluble fluorescent dye into the interior of the core microspheres, the lipid-soluble fluorescent dye may be embedded into the interior of the gel layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by using the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed in the inside of the first coating layer and/or the inside of the gel layer.

Referring to FIG. 8 , a seventh embodiment of the present application proposes a method for making magnetic beads. The method includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by using the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed in the inside of the first coating layer and/or the inside of the gel layer.

S50: Using swelling technology to combine magnetic microspheres and liposoluble fluorescent dyes, the liposoluble fluorescent dyes are dispersed and distributed in the interior of the first coating layer and/or the interior of the gel layer.

Referring to FIG. 9 , an eighth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

S60: The swelling technology is used to combine magnetic microspheres and liposoluble fluorescent dyes, magnetic microspheres and third magnetic nanoparticles, so that the liposoluble fluorescent dyes and the third magnetic nanoparticles are dispersed and distributed inside the first coating layer. / or the inside of the gel layer.

Specifically, magnetic microspheres are dispersed in a first medium, a second medium containing lipid-soluble magnetic nanoparticles is provided, a third medium containing lipid-soluble fluorescent dyes is provided, mixed, and then a swelling reaction (vortex) is performed. Disperse uniformly, rotate for a preset time), liposoluble magnetic nanoparticles and liposoluble fluorescent dyes are embedded in the interior of the core microspheres, magnetically separate the supernatant, and obtain the core microspheres with lipid-soluble magnetic nanoparticles and lipids distributed inside the core microspheres. Fluorescent magnetic beads for soluble fluorescent dyes.

Further, during the process of embedding the lipid-soluble magnetic nanoparticles and the lipid-soluble fluorescent dyes into the core microspheres, the lipid-soluble magnetic nanoparticles and the lipid-soluble fluorescent dyes can be embedded in the interior of the gel layer.

Further, step S60 can be performed multiple times, respectively combining lipid-soluble fluorescent dyes with different fluorescent characteristics and magnetic microspheres, or, respectively combining lipid-soluble fluorescent dyes and magnetic microspheres with different concentrations, and combining lipid-soluble magnetic nanospheres. Particles and magnetic microspheres to obtain multiple magnetic beads with different fluorescence intensities, that is, fluorescently encoded magnetic beads. Among them, different liposoluble fluorescent dyes can be mixed in different proportions to prepare different encoded magnetic beads. It is precisely because of the different ratios of different lipid-soluble fluorescent dyes that the prepared fluorescently encoded magnetic beads have different fluorescent characteristics.

Referring to FIG. 10 , the ninth embodiment of the present application proposes a method for making magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Referring to FIG. 11 , a tenth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Referring to FIG. 12 , an eleventh embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Referring to FIG. 13 , the twelfth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

S50: Using swelling technology to combine magnetic microspheres and liposoluble fluorescent dyes, the liposoluble fluorescent dyes are dispersed and distributed in the interior of the first coating layer and/or the interior of the gel layer.

Referring to FIG. 14 , the thirteenth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S60: The swelling technology is used to combine magnetic microspheres and lipid-soluble fluorescent dyes, magnetic microspheres and third magnetic nanoparticles, so that the lipid-soluble fluorescent dyes and the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Referring to FIG. 15 , the fourteenth embodiment of the present application proposes a method for manufacturing magnetic beads. The method includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Referring to FIG. 16 , a fifteenth embodiment of the present application proposes a method for manufacturing magnetic beads. The method includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Referring to FIG. 17 , the sixteenth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S50: Magnetic microspheres and lipid-soluble fluorescent dyes are combined using swelling technology, so that the lipid-soluble fluorescent dyes are dispersed and distributed inside the first coating layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Referring to FIG. 18 , a seventeenth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S40: The magnetic microspheres and the third magnetic nanoparticles are combined by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

S50: Using swelling technology to combine magnetic microspheres and liposoluble fluorescent dyes, the liposoluble fluorescent dyes are dispersed and distributed in the interior of the first coating layer and/or the interior of the gel layer.

Referring to FIG. 19 , an eighteenth embodiment of the present application proposes a method for manufacturing magnetic beads, which includes the following steps:

S10: Provide a magnetic microsphere, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and a magnetic nanoparticle coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle first cladding.

S20: The magnetic microspheres and the second magnetic nanoparticles are combined by the adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer.

S60: The swelling technology is used to combine magnetic microspheres and lipid-soluble fluorescent dyes, magnetic microspheres and third magnetic nanoparticles, so that the lipid-soluble fluorescent dyes and the third magnetic nanoparticles are dispersed and distributed inside the first coating layer.

S30: Coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and coating at least part of the second magnetic nanoparticles with the gel layer.

Optionally, the gel layer in step S30 of the fourth embodiment to the eighteenth embodiment can be replaced with a second polymer coating layer, and the material of the second polymer coating layer is polystyrene, polymethyl At least one of methyl acrylate, polyethylene-methyl methacrylate, polyacrylonitrile, polyethylene, polypropylene, and polyethyl acrylate.

Thus, the structures of the magnetic beads 10 prepared by the fourth embodiment to the eighteenth embodiment or their alternatives are shown in FIG. 24 or FIG. 25 , and the magnetic bead 10 includes: magnetic microspheres 11 , lipid-soluble fluorescent dyes 15 . The second magnetic nanoparticles 12 , the third magnetic nanoparticles 13 and the second coating layer 14 . Among them, the lipid-soluble fluorescent dye 15 and the third magnetic nanoparticles 13 are dispersed and distributed inside the first coating layer 113. It can be understood that, after entering the interior of the magnetic microspheres 11 through swelling, the lipid-soluble fluorescent dye 15, the third The three magnetic nanoparticles 13 are embedded inside the core microspheres 111 . The second magnetic nanoparticles 12 may be coupled to the outer surface of the core microspheres 111 . The second coating layer 14 coats the outer surface of the magnetic microspheres 11 , and the second coating layer 14 coats at least part of the second magnetic nanoparticles 12 . Optionally, the second coating layer 14 covers all the second magnetic nanoparticles 12, and the outer surface of the magnetic beads 10 is a smooth surface. Wherein, the above-mentioned second coating layer 14 may be a gel layer or a second polymer coating layer. It can be understood that the third magnetic nanoparticles 13 and the liposoluble fluorescent dyes 15 can be dispersed in the gel layer, but limited by the material of the polymer coating layer, the third magnetic nanoparticles 13 and the liposoluble fluorescent dyes 15 is not dispersible in the interior of the second polymer coating.

Further, step S30 in the fourth embodiment to the tenth embodiment above can be deleted, and the structure of the magnetic bead 10 obtained therefrom is shown in FIG. 28 . The magnetic bead 10 includes: Fluorescent dye 15 , second magnetic nanoparticles 12 , and third magnetic nanoparticles 13 . Among them, the lipid-soluble fluorescent dye 15 and the third magnetic nanoparticles 13 are dispersed and distributed inside the first coating layer 113. It can be understood that, after entering the interior of the magnetic microspheres 11 through swelling, the lipid-soluble fluorescent dye 15, the third The three magnetic nanoparticles 13 are embedded inside the core microspheres 111 . The second magnetic nanoparticles 12 may be coupled to the outer surface of the core microspheres 111 .

Referring to FIG. 20, further, in the above-mentioned first embodiment to the eighteenth embodiment, step S30 specifically includes the following steps:

S31: Mix the magnetic microspheres coupled with the second magnetic nanoparticles and the gel material, add a cross-linking agent, and perform a cross-linking reaction, so that the outer surfaces of the magnetic microspheres coupled with the second magnetic nanoparticles are Coat the gel layer.

When the gel material is chitosan, the cross-linking agent can be glutaraldehyde, wherein the mass ratio of chitosan to magnetic microspheres is 0.01-20%, and chitosan interacts with glutaraldehyde through electrostatic interaction. A chemical reaction occurs to produce cross-linking.

Further, the operating temperature of step S20 in the above embodiment is 0-100°C (for example, 0°C, 25°C, 30°C, 50°C, 65°C, 100°C), and the operating temperature of step S30 is 20-65°C ( For example, 20°C, 30°C, 50°C, 65°C), the operating temperature of step S40 is 0～65°C (eg 0°C, 25°C, 30°C, 50°C, 65°C), and the operating temperature of step S50 is 0～65°C 65°C (eg 0°C, 25°C, 30°C, 50°C, 65°C). Within the above temperature range, the increase in autofluorescence of the magnetic microspheres caused by heating can be effectively avoided, thereby improving the detection sensitivity.

Preferably, the operating temperature of step S20 in the above embodiment is 0-30°C (eg 0°C, 25°C, 30°C), and the operating temperature of step S30 is 20-30°C (eg 20°C, 25°C, 30°C) ), the operating temperature of step S40 is 0-30°C (eg 0°C, 25°C, 30°C), and the operating temperature of step S50 is 0-30°C (eg 0°C, 25°C, 30°C). Within the above operating temperature range, the increase in autofluorescence of the magnetic microspheres caused by heating can be effectively avoided, thereby improving the detection sensitivity.

More preferably, the operating temperature of step S20 in the above-mentioned embodiment is room temperature (23 ℃ ± 2 ℃), the operating temperature of step S30 is room temperature (23 ℃ ± 2 ℃), and the operating temperature of step S40 is room temperature (23 ℃ ± 2 ℃). 2° C.), and the operating temperature of step S50 is room temperature (23° C.±2° C.). Within the above operating temperature range, the increase in autofluorescence of the magnetic microspheres caused by heating can be effectively avoided, thereby improving the detection sensitivity.

Different from the situation in the prior art, the magnetic beads of the present application include: magnetic microspheres, second magnetic nanoparticles, a gel layer and third magnetic nanoparticles, and the magnetic microspheres include core microspheres, which are coupled to the core microspheres. The first magnetic nanoparticle on the outer surface of the sphere and the first coating layer coated on the outer surface of the core microsphere and the outer surface of the first magnetic nanoparticle, and the second magnetic nanoparticle is coupled to the outer surface of the magnetic microsphere, condensing The adhesive layer coats the outer surface of the magnetic microspheres and coats at least part of the second magnetic nanoparticles, and the third magnetic nanoparticles are dispersed in the interior of the first coating layer and/or the interior of the gel layer. Under the premise of excessive adsorption of magnetic nanoparticles on the surface of the magnetic microspheres, the magnetic response of the magnetic beads can be improved, the interference signal caused by impurities can be reduced, the autofluorescence of the magnetic microspheres can be reduced, and the detection sensitivity can be improved.

In the several embodiments provided in this application, it should be understood that the disclosed method and device may be implemented in other manners. For example, the device implementations described above are only illustrative. For example, the division of the modules or units is only a logical function division. In actual implementation, there may be other divisions. For example, multiple units or components may be Incorporation may either be integrated into another system, or some features may be omitted, or not implemented.

The units described as separate components may or may not be physically separated, and components displayed as units may or may not be physical units, that is, may be located in one place, or may be distributed to multiple network units. Some or all of the units may be selected according to actual needs to achieve the purpose of the solution in this implementation manner.

In addition, each functional unit in each embodiment of the present application may be integrated into one processing unit, or each unit may exist physically alone, or two or more units may be integrated into one unit. The above-mentioned integrated units may be implemented in the form of hardware, or may be implemented in the form of software functional units.

The above are only the embodiments of the present application, and are not intended to limit the scope of the patent of the present application. Any equivalent structure or equivalent process transformation made by using the contents of the description and drawings of the present application, or directly or indirectly applied to other related technologies Fields are similarly included within the scope of patent protection of this application.

Claims (18)

1. a magnetic bead, is characterized in that, described magnetic bead comprises: Magnetic microspheres, including core microspheres, first magnetic nanoparticles coupled to the outer surface of the core microspheres, and first magnetic nanoparticles coated on the outer surfaces of the core microspheres and the first magnetic nanoparticles cladding; second magnetic nanoparticles coupled to the outer surface of the first coating layer; The third magnetic nanoparticles are dispersed and distributed inside the first coating layer. 2. The magnetic bead according to claim 1, wherein the magnetic bead further comprises: A second cladding layer coats the outer surface of the magnetic microspheres, and the second cladding layer coats at least part of the second magnetic nanoparticles. 3. The magnetic bead according to claim 2, characterized in that, The second coating layer is a gel layer or a second polymer coating layer; Wherein, the third magnetic nanoparticles are dispersed and distributed inside the gel layer. 4. The magnetic bead according to claim 3, characterized in that, The material of the gel layer is at least one of chitosan, sodium alginate, polyacrylic acid, polymethacrylic acid, polyacrylamide, and polyN-polyacrylamide; The material of the polymer first coating layer is at least one of polystyrene, polymethyl methacrylate, polyethylene-methyl methacrylate, polyacrylonitrile, polyethylene, polypropylene, and polyethyl acrylate. kind. 5 . The magnetic bead according to claim 1 , wherein the core microspheres are silica core microspheres, and the first coating layer is a first polymer coating layer. 6 . 6. The magnetic bead according to claim 1, wherein the outer surface of the core microsphere contains a first charged functional group, and the core microsphere is coupled to the first charged functional group through the first charged functional group Magnetic nanoparticles. 7 . The magnetic bead according to claim 1 , wherein the outer surface of the first coating layer contains a second charged functional group, and the first coating layer is coupled through the second charged functional group. 8 . the second magnetic nanoparticles. 8. The magnetic bead according to claim 1, wherein, The first magnetic nanoparticles are water-soluble magnetic nanoparticles; The second magnetic nanoparticles are water-soluble magnetic nanoparticles; The third magnetic nanoparticles are fat-soluble magnetic nanoparticles. 9. The magnetic bead according to claim 8, characterized in that, The fat-soluble magnetic nanoparticles are nanoparticles with paramagnetic properties, and the fat-soluble magnetic nanoparticles contain at least one fat-soluble ligand of unsaturated fatty acid, saturated fatty acid, unsaturated fatty amine or saturated fatty amine. 10. The magnetic bead according to claim 8, characterized in that, The water-soluble magnetic nanoparticles are paramagnetic nanoparticles. 11. The magnetic bead according to claim 3, wherein the magnetic bead further comprises: The fat-soluble fluorescent dye is dispersed and distributed at least inside the first coating layer. 12. The magnetic bead according to claim 1, wherein, In terms of mass percentage, the magnetic beads include: 50%-99.5% of the magnetic microspheres, 0.1%-49.9% of the second magnetic nanoparticles, and 0.1%-49.9% of the third magnetic nanoparticles. 13. A method for making magnetic beads, wherein the method comprises: A magnetic microsphere is provided, the magnetic microsphere includes a core microsphere, a first magnetic nanoparticle coupled to the outer surface of the core microsphere, and the outer surface of the core microsphere and the first magnetic nanoparticles a first coating layer on the outer surface of the magnetic nanoparticle; Using adsorption technology to combine the magnetic microspheres and the second magnetic nanoparticles, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer; Wherein, before the step of combining the magnetic microspheres and the second magnetic nanoparticles by the adsorption technique, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer, or, in After the step of combining the magnetic microspheres and the second magnetic nanoparticles using adsorption technology, so that the second magnetic nanoparticles are coupled to the outer surface of the first coating layer, the method further includes: The magnetic microspheres and the third magnetic nanoparticles are combined by a swelling technique, so that the third magnetic nanoparticles are dispersed and distributed in the inside of the first coating layer and/or the inside of the gel layer. 14 . The method according to claim 13 , wherein the magnetic microspheres and the second magnetic nanoparticles are combined using an adsorption technique, so that the second magnetic nanoparticles are coupled to the first magnetic nanoparticles. 15 . After the step of coating the outer surface of the layer, the method further includes: coating a second coating layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and the first The second coating layer covers at least part of the second magnetic nanoparticles. 15 . The method according to claim 13 , wherein the magnetic microspheres and the second magnetic nanoparticles are combined using an adsorption technique, so that the second magnetic nanoparticles are coupled to the first magnetic nanoparticles. 16 . After the step of coating the outer surface of the layer, the method further includes: coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and the gel layer At least a portion of the second magnetic nanoparticles are coated. 16. The method of claim 13, wherein the step of providing a magnetic microsphere comprises: Combining the core microspheres and the first magnetic nanoparticles using adsorption technology, so that the first magnetic nanoparticles are coupled to the outer surface of the core microspheres; A first coating layer is coated on the outer surface of the core microspheres coupled with the first magnetic nanoparticles, and the first coating layer coats at least part of the first magnetic nanoparticles. 17. The method of claim 15, wherein: The adsorption technology is used to combine the magnetic microspheres and the second magnetic nanoparticles, so that the operating temperature of the second magnetic nanoparticles coupled to the outer surface of the first coating layer is 0-100°C; the operating temperature of coating a gel layer on the outer surface of the magnetic microspheres coupled with the second magnetic nanoparticles, and the gel layer coating at least part of the second magnetic nanoparticles 20～65℃; The operation of combining the magnetic microspheres and the third magnetic nanoparticles by the swelling technology, so that the third magnetic nanoparticles are dispersed and distributed in the interior of the magnetic microspheres and/or the interior of the gel layer The temperature is 0 to 65°C. 18. The method of claim 15, wherein the method further comprises: The magnetic microspheres and the liposoluble fluorescent dye are combined by swelling technology, so that the liposoluble fluorescent dye is dispersed and distributed inside the first coating layer.

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