JP6769728B2 - Fluorescent particles and method for producing fluorescent particles - Google Patents

Description

The present invention relates to fluorescent particles and a method for producing fluorescent particles.

Compared to inorganic phosphors, organic phosphors have the advantages of lower manufacturing costs and better dispersibility in media due to their lower specific gravity, so dye lasers, bioimaging, luminescent dyes for organic EL, and light for solar cells. It is used in a wide range of fields such as wavelength conversion materials.

However, as a characteristic of organic phosphors, it is known that under high concentration conditions, the brightness is significantly reduced due to concentration quenching. As a means for avoiding this concentration quenching, an organic phosphor is used in a diluted state in which a low molecular weight or high molecular weight host material or a solvent is uniformly dispersed at the molecular level.

On the other hand, contrary to the conventional organic phosphors, so-called Aggregation induced emission (AIE) molecules, in which luminescence increases remarkably when aggregated, have been found, which is a problem of conventional organic phosphors. It is expected to overcome the above problems and realize new applications of organic phosphors in the medical and industrial fields. The inventions described in Patent Documents 1 to 3 have been made with respect to such aggregation-induced luminescent molecules.

U.S. Patent Application Publication No. 2014/3287964 U.S. Patent Application Publication No. 2013/089889 U.S. Patent Application Publication No. 2014/017130

An object to be solved by the present invention is to provide novel fluorescent particles stained with aggregation-induced luminescent molecules.

The problem can be solved by the following means.
In the present invention, notations such as "a to b" representing a numerical range are synonymous with a or more and b or less, and a and b are included in the range.

<1> Fluorescent particles comprising swellable polymer particles containing agglutination-induced luminescent molecules.

<2> The fluorescent particle according to <1>, wherein the aggregation-induced luminescent molecule is tetraphenylethylene, a derivative thereof, a siror ring-containing compound or a rhodamine-based compound.

<3> The swellable polymer particles are a polymerizable unsaturated aromatic compound, a polymerizable unsaturated carboxylic acid compound, a polymerizable unsaturated sulfonic acid compound or a salt thereof, a polymerizable carboxylic acid ester compound, or an unsaturated carboxylic acid amide compound. The fluorescent particles according to <1> or <2>, which are particles obtained by using at least one compound selected from the group consisting of a polymerizable unsaturated nitrile compound, a vinyl halide compound, and a conjugated diene compound. ..

<4> The fluorescent particle according to any one of <1> to <3>, which contains the aggregation-induced luminescent molecule in an amount exceeding 0.1 μmol / g per 1 g of the fluorescent particle.

<5> The fluorescent particle according to any one of <1> to <4>, which has a volume average particle diameter of 0.01 to 50 μm.

<6> The fluorescent particle according to any one of <1> to <5>, which is used for labeling.

<7> Produced by a method including a step of swelling the swellable polymer particles in a liquid containing aggregation-induced luminescent molecules and incorporating at least the aggregation-induced luminescent molecules into the swelling polymer particles, <1> to < The fluorescent particle according to any one of 6>.

<8> A method for producing fluorescent particles, which comprises a step of swelling swellable polymer particles in a liquid containing aggregation-induced luminescent molecules and incorporating at least the aggregation-induced luminescent molecules into the swelling polymer particles.

According to the present invention, it is possible to provide novel fluorescent particles stained with aggregation-induced luminescent molecules. According to such fluorescent particles, aggregation-induced luminescent molecules can be made to emit light without agglomerating in a liquid.
Further, the aggregation-induced luminescent molecule does not quench the concentration, and the fluorescent particles having high fluorescence intensity can be obtained also in the present invention. Therefore, by using such fluorescent particles, high-sensitivity detection and diagnosis can be performed, and such fluorescent particles are suitable as labeling particles and the like for next-generation high-sensitivity detection / diagnosis devices and the like. It is thought that it can be used for.

≪Fluorescent particles≫
The fluorescent particles of the present invention are characterized by consisting of swellable polymer particles containing agglutination-induced luminescent molecules.

As a result of diligent research by the present inventor, it is possible to emit light by incorporating aggregation-induced luminescent molecules into predetermined particles and using particles containing the aggregation-induced luminescent molecules, instead of aggregating them in a liquid. We found that we could do it, and further found that the emission intensity increased or decreased by increasing or decreasing the amount of aggregation-induced luminescent molecules incorporated into the particles. That is, it can be said that the present inventor has found a new luminescent means for aggregation-induced luminescent molecules.

When the aggregation-induced luminescent molecules are aggregated in a liquid to emit light, it is not easy to control the aggregation state, for example, the amount of aggregation, and it is easy to control the emission intensity and stably produce the desired emission. It wasn't.
However, according to the present invention, as described above, the aggregation-induced luminescent molecules are not aggregated in the liquid, but are incorporated into predetermined particles to emit light, so that a state in which stable emission can be achieved is maintained. In addition, the emission intensity can be increased or decreased as the amount incorporated into the particles is increased or decreased, so that the emission intensity can be easily controlled.

<Swellable polymer particles>
The swellable polymer particles refer to particles whose volume expands by absorbing the solvent when the polymer particles are immersed in a solvent, and the particles when the polymer particles are immersed in the solvent. Is preferably a particle whose volume expands by about 50 to 800% as compared with that before immersion by absorbing the solvent.

Whether or not the particles are swellable can be confirmed, for example, by observing the particles during solvent absorption with an optical microscope. A method for measuring the amount of swelling will be described by taking polystyrene particles as an example. First, the diameters of any 100 polystyrene particles before swelling are measured by observation with an optical microscope, and the average value of the volume is calculated from the obtained diameters. Next, the polystyrene particles are immersed in a 30% by volume tetrahydrofuran aqueous solution for 5 minutes, and the average value of the volumes of the immersed particles is similarly calculated from the diameters of 100 arbitrary particles by observing with an optical microscope. The average value of the volume of the particles after swelling is calculated with the average value of the volumes of the particles before immersion as 100%. The solvent may be appropriately selected according to the material of the particles by the dyeing method of the present invention.

The polymer particles are not particularly limited, but are a polymerizable unsaturated aromatic compound, a polymerizable unsaturated carboxylic acid compound, a polymerizable unsaturated sulfonic acid compound or a salt thereof, a polymerizable carboxylic acid ester compound, and an unsaturated carboxylic acid amide. It is preferable that the particles are obtained by using at least one compound (A) selected from the group consisting of a compound, a polymerizable unsaturated nitrile compound, a vinyl halide compound, and a conjugated diene compound, and a polymerizable unsaturated fragrance. Particles obtained by using at least one compound selected from the group consisting of a group compound, a polymerizable unsaturated carboxylic acid compound, a polymerizable unsaturated sulfonic acid compound or a salt thereof, and a polymerizable carboxylic acid ester compound. Is more preferable.

Specific examples of such compound (A) include styrene, chlorostyrene, α-methylstyrene, divinylbenzene, vinyltoluene, vinylnaphthalene, divinylnaphthalene, α-naphthyl (meth) acrylate, and (meth) acrylic acid. Polymerizable unsaturated aromatics such as β-naphthyl; polymerizable unsaturated carboxylic acids such as (meth) acrylic acid, itaconic acid, maleic acid, and fumaric acid; polymerizable unsaturated sulfonic acids such as sodium styrene sulfonic acid or its like. Salts; methyl (meth) acrylate, ethyl (meth) acrylate, -n-butyl (meth) acrylate, -2-hydroxyethyl (meth) acrylate, glycidyl (meth) acrylate, ethylene glycol-di-( Polymerizable carboxylic acid esters such as (meth) acrylic acid ester and (meth) acrylic acid tribromophenyl; and the like.

In addition, as compound (A), (meth) acrylonitrile, (meth) acrolein, (meth) acrylamide, N-methylol (meth) acrylamide, methylenebis (meth) acrylamide, butadiene, isoprene, vinyl acetate, vinyl pyridine, N-vinyl Examples thereof include unsaturated carboxylic acid amides such as pyrrolidone, vinyl chloride, vinylidene chloride and vinyl bromide, polymerizable unsaturated nitriles, vinyl halides and conjugated dienes.

The polymer particles may have at least a part of hydrogen contained in the structural unit replaced with a charged substituent or a substituent capable of forming a covalent bond. Such a polymer is synthesized by using a (derivatized) monomer in which at least one hydrogen atom is replaced with the above-mentioned substituent (in this case, an amino group) as a raw material, for example, 4-aminostyrene. Can be done. Further, the polymer may contain a structural unit having a functional group for surface-modifying the obtained fluorescent particles. For example, by using a monomer such as glycidyl methacrylate having an epoxy group as a raw material, fluorescent particles for fluorescent labeling in which the epoxy group is oriented on the surface can be prepared. This epoxy group may be converted to an amino group by reacting with excess aqueous ammonia. Through the amino group thus formed, various biomolecules can be introduced according to a known method (via a molecule that serves as a linker, if necessary).

The polymer particles preferably contain 80% by mass or more of structural units derived from a polymerizable unsaturated aromatic compound and / or a polymerizable carboxylic acid ester compound with respect to 100% by mass of the particles, and 90% by mass or more. Particles are preferable, and particles containing 95% by mass or more are more preferable. Since aromatic compounds have a particularly high affinity for water-insoluble fluorescent dyes, which will be described later, the use of polymer particles having a structural unit derived from aromatic compounds further suppresses the shedding of aggregation-induced luminescent molecules from the fluorescent particles. It is preferable from the viewpoint of being able to do it.

The polymer particles include a polymerizable unsaturated aromatic compound and / or a polymerizable carboxylic acid ester compound, and at least one selected from a polymerizable unsaturated carboxylic acid compound and a polymerizable unsaturated sulfonic acid compound or a salt thereof. The particles of the copolymer obtained by using the raw material are preferable, and the particles of the copolymer of styrene and the polymerizable unsaturated carboxylic acid compound or the polymerizable unsaturated sulfonic acid compound are more preferable.

In the copolymer, a copolymerization of a polymerizable unsaturated aromatic compound and / or a polymerizable carboxylic acid ester compound with at least one selected from a polymerizable unsaturated carboxylic acid compound and a polymerizable unsaturated sulfonic acid compound or a salt thereof. The polymerization ratio is preferably at least one ratio selected from the polymerizable unsaturated carboxylic acid compound and the polymerizable unsaturated sulfonic acid compound or a salt thereof, with the total mass of these being 100% by mass, from the viewpoint of particle dispersibility. Is 0.01 to 5% by mass, more preferably 0.1 to 3% by mass, and even more preferably 0.3 to 1.0% by mass.

In addition to the above-mentioned polymers, the polymer includes, for example, melamine, urea, guanamines (eg, benzoguanamine, acetoguanamine), phenols (eg, phenol, cresol, xylenol), xylene, and derivatives thereof. Examples thereof include other resins containing a structural unit formed from at least one monomer selected from the above. Any one of these monomers may be used alone, or two or more thereof may be used. If desired, one or more comonomer other than the above-mentioned monomers may be used in combination.

Specific examples of other resins include melamine / formaldehyde resin, urea formaldehyde resin, benzoguanamine / formaldehyde resin, phenol / formaldehyde resin, and metaxylene / formaldehyde resin. As the raw material of these resins, not only the monomer itself as described above, but also a prepolymer obtained by reacting the monomer with a compound such as formaldehyde or another cross-linking agent in advance may be used. For example, in the production of melamine / formaldehyde resin, methylol melamine prepared by condensing melamine and formaldehyde under alkaline conditions is generally used as a prepolymer, and the compound is further alkyl etherified (in water). It may be methylated to improve stability in formaldehyde, butylated to improve solubility in an organic solvent, etc.).

Further, the other resin may have at least a part of hydrogen contained in the constituent unit replaced with a charged substituent or a substituent capable of forming a covalent bond. Such a resin can be synthesized by using a monomer in which at least one hydrogen is replaced with the substituent (derivatized) by a known method as a raw material. Melamine resin, urea resin, benzoguanamine resin, etc. usually have an amino group or a cation generated from a site derived from the amino group, and phenol resin, xylene resin, etc. usually have an anion generated from a hydroxyl group or a site derived from the hydroxyl group. Has. Such a resin can be synthesized according to a known method. For example, the melamine / formaldehyde resin can be synthesized by polycondensing the methylolmelamine prepared in advance as described above by heating it after adding a reaction accelerator such as an acid, if necessary.

As the polymer particles, a commercially available product may be used, or those synthesized by a conventionally known method may be used.
The conventionally known method is not particularly limited, and examples thereof include a dispersion polymerization method, a suspension polymerization method, and an emulsion polymerization method using the compound (A), and the emulsion polymerization method is preferable.

The volume average particle diameter of the polymer particles is preferably in the range of 0.01 to 50 μm, more preferably 0.02 to 40 μm, and even more preferably 0.04 to 20 μm.
When the volume average particle diameter is within the above range, the obtained fluorescent particles can be applied to various uses, and can be particularly preferably used as labeling particles, and the obtained fluorescent particles can be used to detect a target substance. And diagnosis can be performed easily and with high sensitivity.
Specifically, the volume average particle size can be measured by a laser diffraction scattered light particle size distribution measuring device, LS13320 type.

The weight average molecular weight of the polymer particles is preferably in the range of 1,000 to 1,000,000, more preferably 5,000 to 800,000, and 10,000 to 600,000. Is even more preferable.

The swellable polymer particles contained in the fluorescent particles of the present invention may be one kind or two or more kinds, but are usually one kind.

<Aggregation-induced luminescent molecule>
The aggregation-induced luminescent molecule (hereinafter, for convenience of explanation, the “aggregation-induced luminescent molecule” is also referred to as a “fluorescent dye”) is not particularly limited as long as it is a compound having a property of emitting fluorescence by aggregating. Of these, water-insoluble compounds are preferable. The water-insoluble means that the amount of the fluorescent dye dissolved in 100 g of water at 25 ° C. is 3 g or less, and the amount of the fluorescent dye dissolved is preferably 2 g or less, more preferably 1 g or less. Examples include compounds. By using such a water-insoluble compound as the fluorescent dye, the colorability to the polymer particles is excellent, and even if the obtained fluorescent particles are dispersed in water, the fluorescent dye is less likely to fall off, and stable fluorescence intensity is maintained for a long period of time. Fluorescent particles shown in the above can be obtained.

The fluorescent dye may be any known as a compound exhibiting aggregation-induced luminescence, and examples thereof include a hydrocarbon aromatic compound, a heteroaromatic compound, a silol ring-containing compound, and a rhodamine compound. Tetraphenylethylene, a derivative thereof, a silol ring-containing compound, or a rhodamine-based compound is preferable from the viewpoint that it can be easily incorporated into the polymer particles.

<Tetraphenylethylene and its derivatives>
The tetraphenylethylene and its derivative refer to a compound having a tetraphenylethylene skeleton, and examples of the tetraphenylethylene or its derivative include a compound represented by the following formula (1). The compound represented by the following formula (1) is not particularly limited as long as it has a tetraphenylethylene skeleton, and various groups can be introduced into R ^{1 to} R ⁴ depending on the desired use.
Among these, tetraphenylethylene is preferable.

In the above formula (1), R ^{1 to} R ⁴ are independently hydrogen atoms, organic groups or organic metal groups, and hydrogen atoms or aromatic ring-containing organic groups are preferable.
In the above equation (1), a to d are independently integers of 1 to 5. When a is 2 or more, the plurality of R ^1s may be the same or different, and the plurality of R ^1s may be bonded to each other to form a ring. When b to d are two or more, or in other formulas of the present invention, when two or more reference numerals Z are present in one formula, they may be the same or different. Further, R ¹ and R ² , R ² and R ⁴ , R ³ and R ⁴ , and R ³ and R ¹ may be combined to form a ring.

In the above formula (1), the following compounds are preferably mentioned as the compound when at least one of R ^{1 to} R ⁴ is a group other than a hydrogen atom.

（Ｒ₁〜Ｒ₃はそれぞれ独立して、水素原子、炭化水素基又は炭化水素基の少なくとも一部がヘテロ原子で置換された基である。）

(R _{1 to} R ₃ are independent groups in which at least a hydrogen atom, a hydrocarbon group, or a hydrocarbon group is substituted with a hetero atom.)

（Ｒ₁〜Ｒ₄はそれぞれ独立して、水素原子、炭化水素基又は炭化水素基の少なくとも一部がヘテロ原子で置換された基である。）

(R _{1 to} R ₄ are independent groups in which at least a hydrogen atom, a hydrocarbon group, or a hydrocarbon group is substituted with a hetero atom.)

（Ｒ₁〜Ｒ₁₀はそれぞれ独立して、水素原子、炭化水素基又は炭化水素基の少なくとも一部がヘテロ原子で置換された基である。）

(R _{1 to} R ₁₀ are independent groups in which at least a hydrogen atom, a hydrocarbon group, or a hydrocarbon group is substituted with a hetero atom.)

（Ｒ₁〜Ｒ₁₆はそれぞれ独立して、水素原子、炭化水素基又は炭化水素基の少なくとも一部がヘテロ原子で置換された基である。）

(R _{1 to} R ₁₆ are independent groups in which at least a hydrogen atom, a hydrocarbon group, or a hydrocarbon group is substituted with a hetero atom.)

（Ｒ₁〜Ｒ₂₁はそれぞれ独立して、水素原子、炭化水素基又は炭化水素基の少なくとも一部がヘテロ原子で置換された基である。）

(R _{1 to} R ₂₁ are independent groups in which at least a hydrogen atom, a hydrocarbon group, or a hydrocarbon group is substituted with a hetero atom.)

（Ｒ₁〜Ｒ₄、Ｒ₆〜Ｒ₁₇はそれぞれ独立して、水素原子、炭化水素基又は炭化水素基の少なくとも一部がヘテロ原子で置換された基である。）

(R _{1 to} R ₄ and R _{6 to} R ₁₇ are independent groups in which at least a hydrogen atom, a hydrocarbon group, or a hydrocarbon group is substituted with a hetero atom.)

As the derivative of tetraphenylethylene, a compound described in JP-A-2014-12654 and US Patent Application Publication No. 2013/177991 may be used.

<Siror ring-containing compound>
The siror ring-containing compound is not particularly limited as long as it is a aggregation-induced luminescent compound having a siror ring, but a compound represented by the following formula (2) is more preferable. The groups represented by the same reference numerals in the following formula (2) may be the same or different, but are preferably the same.

The ^RA is independently a hydrogen atom or a hydrocarbon group having 1 to 12 carbon atoms, preferably a hydrogen atom or a hydrocarbon group having 1 to 6 carbon atoms, and more preferably a hydrogen atom or a hydrocarbon group having 1 to 4 carbon atoms. It is a hydrocarbon group of, more preferably a hydrogen atom.
Independently, a in the formula (2) is an integer of 1 to 5, preferably 1 or 2.

Wherein R ^B is independently an aromatic ring-containing organic group.
Examples of the aromatic ring include a benzene ring, a naphthalene ring, a pyrrole ring, an imidazole ring, an imidazoline ring, a pyrazole ring, a pyridine ring, a pyrazine ring, a furan ring, a thiophene ring, an oxazole ring, and a thiazole ring.
Wherein R ^B preferably contains at least one benzene ring, more preferably a phenyl group.

The ^RC is independently an organic group, preferably an aromatic ring-containing organic group or a hydrocarbon group having 1 to 20 carbon atoms.
As the hydrocarbon group having 1 to 20 carbon atoms, a hydrocarbon group having 1 to 12 carbon atoms is preferable.
The ^RC is preferably a hydrocarbon group having 1 to 12 carbon atoms, and more preferably a phenyl group or an alkyl group having 1 to 12 carbon atoms.

The organic group in R ^B and R ^C may be a hydrocarbon radical consisting of carbon and hydrogen atoms, nitrogen atom, oxygen atom, or may be a group containing a hetero atom such as sulfur or silicon atom ..

Since the compound represented by the formula (2) has two benzene rings bonded to the siror ring and further has at least two aromatic ring-containing organic groups, the compound is aggregated to suppress intramolecular rotation. , It is considered that light emission occurs. That is, the compound represented by the formula (2) is not particularly limited as long as it has two benzene rings bonded to the siror ring and further has at least two aromatic ring-containing organic groups, and can be used for desired purposes. Depending on the situation, various groups can be introduced.

Examples of the compound represented by the formula (2) include the following compounds.

＊表中、Ｐｈは、フェニル基を示し、Ｍｅはメチル基をし、Ｒは糖鎖を示す。該糖鎖としては、例えば、ラクトース誘導体が挙げられる。

* In the table, Ph indicates a phenyl group, Me indicates a methyl group, and R indicates a sugar chain. Examples of the sugar chain include lactose derivatives.

The compound represented by the formula (2) is, for example, when the R ^C is a group represented by _{-C 6 H 4 -CH 2 -N (} C 2 H 5) 2, in the liquid secondary, The compound represented by the above formula (2) is such that the compound represented by the above formula (2) can have a structure in which the portion is a group represented by −C ₆ H ₄ −CH ₂ −N ⁺ (C ₂ H ₅ ) _2. Also includes cations and the like.

<Rhodamine compounds>
The rhodamine-based compound is not particularly limited as long as it is a aggregation-induced luminescent compound having a skeleton represented by the following formula (3), but an aminobenzopyroxanthene-based dye is preferable and represented by the following formula (3'). The compound to be used is more preferable.
The compound represented by the following formula (3') is not particularly limited as long as it has a specific skeleton, and various groups can be introduced into ^Ra and R ^b depending on the desired use.
The groups represented by the same reference numerals in the following formula (3') may be the same or different, but are preferably the same.

Each of ^Ra and R ^b is independently a hydrogen atom or an organic group having 1 to 20 carbon atoms, preferably a hydrocarbon group having 1 to 12 carbon atoms.

The R ^c and R ^d are independently hydrogen atoms, halogen atoms, or organic groups having 1 to 20 carbon atoms.
The R ^c and R ^d are preferably hydrogen atoms.

Wherein R ^e is independently -COOR ^h or amino group. The R ^h is a hydrogen atom or an organic group having 1 to 20 carbon atoms.
The R ^e is preferably a carboxyl group.

The R ^f is independently an amide bond-containing group which may have a hydrogen atom, a halogen atom, an amino group and a protective group, or an organic group having 1 to 20 carbon atoms, and is preferably a hydrogen atom.

The R ^g is independently a hydrogen atom, a halogen atom, a nitro group, a carboxyl group, an amino group or an organic group having 1 to 20 carbon atoms, and is preferably a hydrogen atom.
m is an integer of 1 to 4, preferably 1.

As the organic group in R ^a , R ^b , R ^c , R ^d , R ^f , R ^g and R ^h , a hydrocarbon group having 1 to 20 carbon atoms and a part of the hydrocarbon group are preferably nitrogen atoms. Examples thereof include a group substituted with an oxygen atom or a sulfur atom, more preferably a hydrocarbon group having 1 to 12 carbon atoms.

The R ^a and R ^c and / or R ^b and R ^d may be bonded to each other to form a heterocycle having 5 or 6 constituent atoms containing at least one nitrogen atom. Specific examples thereof include compounds represented by the following formulas (3-1) and (3-2).
Examples of the heterocycle include pyrrolidine, pyrrole, imidazole, pyrazole, piperidine, pyridine, piperazine, pyridazine, pyrimidine and pyrazine. Of these, piperidine is preferred.
The heterocycle may have a substituent R ^j . The R ^j is independently a hydrocarbon group having 1 to 12 carbon atoms or a group in which a part of the hydrocarbon group is substituted with a nitrogen atom, an oxygen atom or a sulfur atom.

［式（３−１）及び（３−２）中、Ｒ^a、Ｒ^c、Ｒ^e、Ｒ^f、Ｒ^g及びｍはそれぞれ独立に、式（３'）中のＲ^a、Ｒ^c、Ｒ^e、Ｒ^f、Ｒ^g及びｍと同義である。ｎは独立に０〜６の整数であり、好ましくは０〜２の整数である。］

[In equations (3-1) and (3-2), R ^a , R ^c , R ^e , R ^f , R ^g and m are independent of each other, and R ^a , R ^c and R in equation (3'), respectively. Synonymous with ^e , R ^f , R ^g and m. n is independently an integer of 0 to 6, preferably an integer of 0 to 2. ]

Further, the R ^e and R ^f may be bonded to each other to form a heterocycle having 5 or 6 constituent atoms.
Examples of the heterocycle include γ-butyrolactone, β-lactam, and γ-lactam. Of these, γ-butyrolactone is preferred.
The heterocycle may have a substituent. Examples of the substituent include a hydrocarbon group having 1 to 12 carbon atoms, a group in which a part of the hydrocarbon group is substituted with a nitrogen atom, an oxygen atom or a sulfur atom, a hydroxyl group, an amino group, a pyridinyl group, and a frill. Examples include a group and a thienyl group.
It is preferable that R ^e and R ^f are bonded to each other to form γ-butyrolactone. Specific examples of such a compound include a compound represented by the following formula (3-3).

［式（３−３）中、Ｒ^a〜Ｒ^d、Ｒ^g及びｍはそれぞれ独立に、式（３'）中のＲ^a〜Ｒ^d、Ｒ^g及びｍと同義である。］

[In equation (3-3), R ^{a to} R ^d , R ^g and m are independently synonymous with R ^{a to} R ^d , R ^g and m in equation (3'). ]

The compound represented by the formula (3') shall represent the structure not only by the description method as described in the formula (3') but also by the description method having a resonance structure as described in the following formula (3A). Can be done. Unless otherwise specified in the present invention, the structure of a rhodamine-based compound is represented by a description method as described in the above formula (3').
Further, since the compound represented by the formula (3') can have a structure as shown in the following formula (3B) in a liquid or the like, the compound represented by the formula (3') is described in the following formula (3B). ) Is also included.

Commercially available products may be used as the fluorescent dye, and conventionally known methods such as US Patent Application Publication No. 2012/299474, US Patent Application Publication No. 2013/177991, US Patent Application Publication No. Compounds synthesized by the methods described in 2013/89889, S. Kamino, et. Al., Chem. Commun., 2010, 46, 9013-9015 may be used.

As the fluorescent dye, a dye capable of retaining a sufficient amount in the polymer particles in the infiltration method described later is preferable, and the dye has high solubility in an organic solvent (eg, THF) used in the infiltration method described below. More preferably, a dye having high compatibility with the polymer particles and such that the fluorescent dye does not leak from the particles even if the obtained fluorescent particles are dispersed in an aqueous medium is particularly preferable. When a fluorescent dye having poor compatibility with (the polymer constituting the polymer) and having high hydrophilicity is used and the obtained fluorescent particles are dispersed in an aqueous medium, the fluorescent dye fluoresces. There is a risk of leakage from the particles.

The fluorescent particles of the present invention preferably contain a fluorescent dye in an amount of more than 0.1 μmol / g, more preferably 0.2 μmol / g or more, still more preferably 20 μmol / g or more per 1 g of fluorescent particles. .. When the amount of the fluorescent dye in the fluorescent particles is equal to or more than the above range, a sufficient amount of fluorescence intensity is exhibited, which is preferable.

In the present invention, the amount of the fluorescent dye introduced into the polymer particles is determined by quantitatively analyzing the amount of the fluorescent dye after thermal decomposition by pyrolysis gas chromatography-mass spectrum (hereinafter, also referred to as "PyGC-MS"). Can be sought. Specifically, the fluorescent particles are dissolved in a solvent that dissolves the particles, for example, chloroform, methylene chloride, toluene, xylene, N-methylpyrrolidone, or the like to prepare a uniform solution having a constant concentration. The amount of the fluorescent dye introduced can be calculated by driving and analyzing a fixed amount of the obtained solution by PyGC-MS.

The fluorescent dye contained in the fluorescent particles of the present invention may be one kind or two or more kinds.

<Physical properties of fluorescent particles, etc.>
The volume average particle diameter of the fluorescent particles of the present invention is preferably in the range of 0.01 to 50 μm, more preferably 0.02 to 40 μm, and even more preferably 0.04 to 20 μm.
When the volume average particle diameter is in the above range, the fluorescent particles can be applied to various uses, and can be particularly preferably used as labeling particles, and the fluorescent particles make it easy to detect and diagnose the target substance. In addition, it can be performed with high sensitivity.
Specifically, the volume average particle size can be measured by a laser diffraction scattered light particle size distribution measuring device, LS13320 type.

The degree of dispersion of the particle size of the fluorescent particles of the present invention is preferably 15% or less, more preferably 12% or less, further preferably 10% or less.
When the degree of dispersion is within the above range, there is no variation in fluorescence intensity, and high reproducibility can be ensured.
Specifically, the degree of dispersion can be measured by a laser diffraction scattered light particle size distribution measuring device, LS13320 type.

The fluorescent particles of the present invention preferably have an average specific gravity in the range of 0.90 to 1.5 from the viewpoint of storage stability and the like.
The average specific gravity is measured as follows.
Prepare solutions of various specific gravities measured using a hydrometer, float the powder of the dried fluorescent particle group on this solution, observe the sedimentation state of the powder, and the powder does not completely sink. The specific gravity of the solution used when floating in the solution is defined as the average specific gravity in the present invention.

<Use of fluorescent particles, etc.>
As described above, the fluorescent particles of the present invention can maintain a state in which they can emit light stably, and the emission intensity can be easily controlled. Therefore, various fields using fluorescence (dye laser, biotechnology). It can be applied to a wide range of fields such as imaging, luminescent dyes for organic EL, and optical wavelength conversion materials for solar cells). In particular, according to the present invention, since fluorescent particles having high luminescence intensity can be obtained, when used as labeling particles such as an immunoassay reagent for determining the presence of a substance to be detected by luminescence, it is visually observed. Judgability and detectability by a fluorescence detector are significantly improved.
As an example of this application, biology such as enzyme immunoassay, fluorescence immunoassay, latex agglutination, immunochromatography, etc. using antigen-antibody reaction by binding an antigen (or antibody) to the surface of fluorescent particles. Various methods using the target reaction can be mentioned.

≪Manufacturing method of fluorescent particles≫
Generally, as a method for producing fluorescent particles, (method 1) a method of immobilizing a fluorescent dye on the surface of the particles by chemical bonding, and (method 2) granulating polymer particles in the presence of the fluorescent dye to form particles. Examples thereof include a method of incorporating the fluorescent dye into the particles in the process of swelling, and an infiltration method including (method 3) a step of swelling the polymer particles in a liquid containing the fluorescent dye and incorporating at least the fluorescent dye into the particles. In the present invention, it is preferable to produce fluorescent particles by (Method 3).

In the above (method 1), since the fluorescent dye is bonded to the particle surface by a chemical bond, the dye is less likely to fall off and stable fluorescence intensity can be obtained, while the amount of the fluorescent dye introduced is small, resulting in high fluorescence intensity fluorescence. There is a tendency that particles cannot be obtained. In (Method 2), it tends to be difficult to produce fluorescent particles having sufficiently high fluorescence intensity, and it is difficult to control the amount of fluorescent dye introduced in the granulation process, resulting in fluorescence. The calibration accuracy of particles tends to decrease.
On the other hand, according to the above (method 3), the amount of the fluorescent dye in the obtained fluorescent particles can be easily controlled to a desired amount, and fluorescent particles having a desired emission intensity can be easily produced.

Hereinafter, a preferable example of the method for producing the fluorescent particles of the present invention will be given.

First, polymer particles made of, for example, a styrene-methacrylic acid copolymer are produced by emulsion polymerization or the like. The preferable composition, particle size and the like are as described above.

Next, this polymer particle group is dispersed in an aqueous medium. The concentration of the polymer particles in the aqueous medium is preferably 0.1 to 20% by mass, more preferably 0.1 to 10% by mass, further preferably 0.2 to 8% by mass, and 0.3 to 5% by mass. % Is particularly preferable.

As the aqueous medium in which the polymer particles are dispersed, a medium containing 50% by mass or more of water is preferable, a medium containing 80% by mass or more is more preferable, and a medium containing 90% by mass or more is further preferable.

The aqueous medium may contain additives such as surfactants.
Examples of the surfactant include nonionic surfactants. Among them, polymer-based surfactants are preferable because they enhance particle dispersion stability, and Kolliphor P407 (manufactured by Sigma-Aldrich) is more preferable. The concentration of the surfactant is preferably contained in the aqueous medium in the range of 0.1 to 1% by mass.

A dye solution in which a fluorescent dye is dissolved in a solvent is added to the aqueous dispersion of the obtained polymer particles, and the mixture is stirred. At this time, it is preferable to heat or heat. When the dye solution is added to the aqueous dispersion of the polymer particles, the polymer particles swell and the fluorescent dye present in the solvent is gradually absorbed into the particles.

Examples of the solvent for dissolving the fluorescent dye include ethers such as ethyl ether and isopropyl ether; alcohols such as methanol and ethanol; methylene chloride; ethylene dichloride; chloroform; carbon tetrachloride; ethyl acetate; methyl acetate; methyl ethyl ketone; Cyclohexane; cyclopentane; tetrahydrofuran (THF); toluene; hexane; heptane and the like can be mentioned, and one or more of them may be used depending on the type of fluorescent dye used. In the present invention, THF is preferable because it is soluble in water and has high solubility of the fluorescent dye.

The concentration of the fluorescent dye in the dye solution is preferably in the range of 0.01 to 20 mmol / L, more preferably 0.1 to 15 mmol / L, still more preferably 1 to 10 mmol / L.

Regarding the ratio of the aqueous dispersion of the polymer particles to the dye solution, the ratio of the polymer particles to the fluorescent dye is preferably in the range of 0.001 to 100 g of the fluorescent dye with respect to 100 g of the polymer particles, and 0.005 to 0.005. 80 g is more preferable, and 0.01 to 50 g is further preferable.

When the fluorescent dye is incorporated into the polymer particles, an organic solvent (solvent that dissolves the fluorescent dye) is preferably present, and the concentration of the organic solvent is preferably 15 to 50% by volume, preferably 20 to 45% by volume. % Is more preferred, and 25-40% by volume is even more preferred. When the concentration of the organic solvent is within the above range, the polymer particles are sufficiently swelled, and the fluorescent dye can be easily incorporated into the polymer particles. In addition, leaching of the fluorescent dye is reduced, and fluorescent particles capable of suppressing aggregation and shape change due to a change in the state of the particles can be easily obtained.

The heating or heating temperature is preferably set in the range of 20 to 30 ° C., which does not exceed the reaction temperature when polymerizing the polymer particles, but usually, the higher the temperature, the higher the uptake rate of the fluorescent dye. It will be faster.

The time for the fluorescent dye to be absorbed by the polymer particles (the time for heating or heating and stirring the aqueous dispersion of the polymer particles and the dye solution as necessary) includes the type of fluorescent dye used, the composition of the polymer particles, the particle size, etc. Although it is determined by, usually 2 to 30 minutes is preferable.

The step of absorbing the fluorescent dye into the polymer particles is completed by removing the solvent used in the dye solution.
Examples of the method for removing the solvent include centrifugation or dialysis. Further, it is preferable to provide a cleaning step with a surfactant. The fluorescent particles treated in this way are preferable because the fluorescent dye does not easily leach out in the aqueous solution.

As described above, the fluorescent dye is absorbed for a predetermined time, the dye solution and the fluorescent particles are separated by centrifugation or the like, and then the fluorescent particles are dispersed in a solvent such as water to obtain a fluorescent particle dispersion.

Hereinafter, the present invention will be described in detail with reference to Examples and Comparative Examples, but the present invention is not limited to these Examples.

[Example 1]
Polystyrene (PS) particle dispersion [solid content 5.2% by mass, volume average particle diameter of polystyrene particles 0.12 μm, dispersion medium: water] 96 μL, 100 μL of water, nonionic surfactant "Kolliphor P407" (Sigma) 50 μL of 2% aqueous solution (manufactured by Aldrich) and 100 μL of 0.01 mmol / L THF solution of “Tetrafeneylylene (hereinafter also referred to as“ TPE ”)” (manufactured by Tokyo Kasei Kogyo Co., Ltd.), which is a coagulation-induced luminescent substance, are added to polystyrene particles. A mixed solution with TPE was prepared. The mixed solution was stirred at 25 ° C. for 2 minutes to prepare polystyrene particles containing TPE.

Using the obtained dispersion of polystyrene particles containing TPE, the particles were precipitated by a centrifugal purification method, the supernatant was removed, and then pure water was added to redisperse the particles. This operation (centrifugal purification and redispersion) was repeated 4 times to obtain a dispersion containing polystyrene particles containing TPE.

A part of this dispersion was sampled, dried at 50 ° C. for 1 hour, and then the surface condition of the obtained particles was confirmed by a scanning electron microscope (hereinafter referred to as "SEM"). As a result of observation, no aggregates of TPE could be confirmed, and it was confirmed that all the charged TPE was incorporated into the polystyrene particles in the obtained particles.

[Example 2]
Same as in Example 1 except that the THF solution concentration of TPE (manufactured by Tokyo Chemical Industry Co., Ltd.) was 1 mmol / L, the addition amount was 10 μL, and 90 μL of THF was added to the mixed solution of polystyrene particles and TPE. By the method, a dispersion containing polystyrene particles containing TPE was obtained. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged TPE was incorporated into the polystyrene particles in the obtained particles.

[Example 3]
Implemented except that "1,1,2,3,4,5-Hexaphenyl polystyrene-2,4-diene (hereinafter also referred to as" HPS ")" (manufactured by Tokyo Chemical Industry Co., Ltd.) was used as the aggregation-induced luminescent substance. A dispersion containing polystyrene particles containing HPS was obtained by the same method as in Example 2. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged HPS was incorporated into the polystyrene particles in the obtained particles.

[Example 4]
<Synthesis of aggregation-induced luminescent rhodamine>
3-Diethylaminoanisole (1 equiv.) Was slowly added dropwise to a methylene chloride solution of phthalic anhydride (1 equiv.) And aluminum chloride (1.1 equiv.), And the mixture was stirred at 0 ° C. for 4 hours under a nitrogen atmosphere. The reaction solution was poured into a 6 mol / L hydrochloric acid aqueous solution, stirred for 10 minutes, and then the organic layer was extracted with methylene chloride. The obtained organic layer was dried over magnesium sulfate, and then the solvent was distilled off under reduced pressure to obtain a crude product. For further purification, the crude product was recrystallized from a mixed solvent of methanol / water to obtain yellow crystals of 2- [4- (Diethylamino) -2-Methoxybenzoyl] benzoic acid (hereinafter, also referred to as “Compound A”). It was.

A methylene chloride solution of boron tribromide (1.9 equiv.) Was added dropwise at −78 ° C. to a methylene chloride solution of compound A (1 quiv.), Stirred for 1 hour, and then the temperature was raised to -25 ° C. After completion of the reaction, quenching with water, distilling off the solvent under reduced pressure, and recrystallization with a mixed solvent of methanol / water were carried out to obtain 2- [4- (Diethylamino) -2-hydroxybenzoyl] benzoic acid (hereinafter referred to as "Compound B"). (Also known as) yellow crystals were obtained.

Compound B (2 equiv.) And resorcinol (1 equiv.) Were added to methanesulfonic acid (2 mL) and heated at 90 ° C. for 2 hours in a sealed tube. The reaction solution was poured into ice water with stirring, and the pH was adjusted to 11 to 12 with a 1 M aqueous sodium hydroxide solution. The obtained solution was stirred for 20 minutes, extracted three times with methylene chloride, the obtained organic layer was dried over magnesium sulfate, and then distilled off under reduced pressure to obtain a crude product. The obtained crude product was purified by silica gel chromatography, further crystallized with acetonitrile, and recrystallized from a rhodamine compound, 3', 3''-Bis (oxospirroisobenzofuran) -3,7-bis (diethylamino) benzopyranoxane (the above formula (the above formula). A compound in which R ^a and R ^b in 3-3) are ethyl groups, R ^c and R ^d are hydrogen atoms, and R ^g is a hydrogen atom; hereinafter also referred to as “BPX”) was obtained. The structure of the obtained compound was confirmed by ¹ H-NMR, ¹³ C-NMR, and MS.

<Preparation of rhodamine-containing particles>
A dispersion containing polystyrene particles containing BPX was obtained by the same method as in Example 2 except that BPX was used as the aggregation-induced luminescent substance. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged BPX was incorporated into the polystyrene particles in the obtained particles.

[Example 5]
A dispersion containing polystyrene particles containing TPE was obtained by the same method as in Example 1 except that the THF solution concentration of TPE (manufactured by Tokyo Chemical Industry Co., Ltd.) was set to 1 mmol / L. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged TPE was incorporated into the polystyrene particles in the obtained particles.

[Example 6]
Instead of polystyrene particle dispersion, polymethylmethacrylate (PMMA) particle dispersion [solid content 15.1% by mass, volume average particle diameter of polymethylmethacrylate particles 0.12 μm, dispersion medium: water] 33 μL was used, and 66 μL of water was used. A dispersion containing polymethylmethacrylate particles containing TPE was obtained by the same method as in Example 1 except that the THF solution concentration of TPE (manufactured by Tokyo Kasei Kogyo Co., Ltd.) was set to 1 mmol / L. As in Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged TPE was incorporated into the polymethylmethacrylate particles in the obtained particles.

[Example 7]
A dispersion containing polystyrene particles containing HPS was obtained by the same method as in Example 5 except that HPS (manufactured by Tokyo Chemical Industry Co., Ltd.) was used as the aggregation-induced luminescent substance. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged HPS was incorporated into the polystyrene particles in the obtained particles.

[Example 8]
A dispersion containing polystyrene particles containing BPX was obtained by the same method as in Example 5 except that BPX was used as the aggregation-induced luminescent substance. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged BPX was incorporated into the polystyrene particles in the obtained particles.

[Example 9]
A dispersion containing polystyrene particles containing TPE was obtained by the same method as in Example 1 except that the THF solution concentration of TPE (manufactured by Tokyo Chemical Industry Co., Ltd.) was set to 10 mmol / L. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged TPE was incorporated into the polystyrene particles in the obtained particles.

[Example 10]
Polystyrene particle dispersion [solid content 5.2% by mass, volume average particle diameter of polystyrene particles 0.12 μm, dispersion medium: water] is 19 μL, and the THF solution concentration of TPE (manufactured by Tokyo Kasei Kogyo Co., Ltd.) is 10 mmol / L. A dispersion containing polystyrene particles containing TPE was obtained by the same method as in Example 1. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged TPE was incorporated into the polystyrene particles in the obtained particles.

[Example 11]
A dispersion containing polystyrene particles containing HPS was obtained by the same method as in Example 10 except that HPS (manufactured by Tokyo Chemical Industry Co., Ltd.) was used as the aggregation-induced luminescent substance. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged HPS was incorporated into the polystyrene particles in the obtained particles.

[Example 12]
A dispersion containing polystyrene particles containing BPX was obtained by the same method as in Example 10 except that BPX was used as the aggregation-induced luminescent substance. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged BPX was incorporated into the polystyrene particles in the obtained particles.

[Example 13]
100 parts by mass of water, 0.1 parts by mass of sodium dodecylbenzenesulfonate and 20 parts by mass of styrene are placed in a 1 L four-necked flask equipped with a cooling tube, a temperature controller and a stirrer, and at 200 rpm. While stirring, 1 part by mass of potassium persulfate dissolved in 50 parts by mass of water was added. After the reaction was carried out for 2 hours after the temperature reached 75 ° C., 80 parts by mass of styrene, 30 parts by mass of TPE (manufactured by Tokyo Kasei Kogyo Co., Ltd.), 0.6 parts by mass of sodium dodecylbenzene sulfonate, and water. A monomer emulsion consisting of 200 parts by mass was added dropwise over 5 hours, and polymerization was continued for 4 hours. An aqueous dispersion of styrene polymer particles containing TPE [solid content 25.4%, volume average particle diameter of particles] 0.15 μm] was obtained. When the particles obtained by SEM were observed in the same manner as in Example 1, it was found that some TPEs were not incorporated outside the particles.

[Comparative Example 1]
In Example 1, 100 μL of a 0.01 mmol / L THF solution of Nile Red (manufactured by Tokyo Chemical Industry Co., Ltd.), which is a fluorescent substance, was used instead of the TPE solution, in the same manner as in Example 1. A dispersion containing polystyrene particles containing Nile Red was obtained.
Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged Nile Red was incorporated into the polystyrene particles in the obtained particles.

[Comparative Example 2]
In Example 2, Nile Red was contained in the same manner as in Example 2 except that 10 μL of a 1 mmol / L THF solution of Nile Red (manufactured by Tokyo Chemical Industry Co., Ltd.) was used instead of the TPE solution. A dispersion containing polystyrene particles was obtained. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged Nile Red was incorporated into the polystyrene particles in the obtained particles.

[Comparative Example 3]
A dispersion containing polystyrene particles containing Nile Red was obtained by the same method as in Comparative Example 1 except that the THF solution concentration of Nile Red (manufactured by Tokyo Chemical Industry Co., Ltd.) was set to 1 mmol / L. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged Nile Red was incorporated into the polystyrene particles in the obtained particles.

[Comparative Example 4]
A dispersion containing polystyrene particles containing Nile Red was obtained by the same method as in Comparative Example 1 except that the THF solution concentration of Nile Red (manufactured by Tokyo Chemical Industry Co., Ltd.) was set to 10 mmol / L. Similar to Example 1, the particles obtained by SEM were observed, and it was confirmed that all the charged Nile Red was incorporated into the polystyrene particles in the obtained particles.

[Comparative Example 5]
In Example 10, Nile Red was contained by the same method as in Example 10 except that the THF solution concentration of Nile Red (manufactured by Tokyo Chemical Industry Co., Ltd.) was 10 mmol / L instead of the TPE solution. A dispersion containing the polystyrene particles was obtained. As a result of observing the particles obtained by SEM in the same manner as in Example 1, agglomerates could be confirmed outside the obtained particles, and in the obtained particles, a part of the charged Nile Red was made of polystyrene particles. It turned out to be outside.

[Confirmation of emission intensity]
The agglomeration-induced luminescent substance or fluorescent substance-containing particle dispersion prepared in the above Examples and Comparative Examples is irradiated with ultraviolet rays using an ultraviolet irradiator (manufactured by Atto Co., Ltd., TP-15MP), and the emission intensity is visually confirmed. Was done. The results are shown in Table 2. The evaluation criteria in Table 2 are as follows.
◎: Very strong ○: Strong △: Slightly strong ×: Weak

Claims (5)

Fluorescent particles composed of swellable polymer particles containing aggregation-induced luminescent molecules .
The aggregation-induced luminescent molecule is at least one compound selected from the compounds represented by the following formulas (1), (2) and (3').
At least one of the swellable polymer particles selected from the group consisting of a polymerizable unsaturated aromatic compound, a polymerizable unsaturated carboxylic acid compound, a polymerizable unsaturated sulfonic acid compound or a salt thereof, and a polymerizable carboxylic acid ester compound. Particles obtained using seed compounds,
Fluorescent particles.

[In the formula (1), R ^{1 to} R ⁴ are independently hydrogen atoms, organic groups or organometallic groups, and a to d are independently integers of 1 to 5, and a plurality of Rs are used. ¹ , a plurality of R ² , a plurality of R ³ or a plurality of R ⁴ may be bonded to each other to form a ring, and R ¹ and R ² , R ² and R ⁴ , R ³ and R ⁴ , R ³ And R ¹ may be combined to form a ring. ]

[In formula (2), ^RA is independently a hydrogen atom or a hydrocarbon group having 1 to 12 carbon atoms, a is independently an integer of 1 to 5, and R ^B is independently containing an aromatic ring. It is an organic group and ^RC is an independent organic group. ]

[In formula (3'), ^Ra and R ^b are independently hydrogen atoms or organic groups having 1 to 20 carbon atoms, and R ^c and R ^d are independently hydrogen atoms, halogen atoms or carbon atoms, respectively. 20 is an organic group, ^{R e} is independently -COOR ^h or amino group ^{(R h} is a hydrogen atom or an organic group having 1 to 20 carbon atoms) is, ^{R f} is independently a hydrogen atom, It is an amide bond-containing group or an organic group having 1 to 20 carbon atoms which may have a halogen atom, an amino group and a protective group, and R ^g is independently a hydrogen atom, a halogen atom, a nitro group, a carboxyl group and an amino. It is a group or an organic group having 1 to 20 carbon atoms, m is an integer of 1 to 4, and R ^a and R ^c and / or R ^b and R ^d are bonded to each other and have a substituent R ^j . It may be a heterocycle having 5 or 6 constituent atoms containing at least one nitrogen atom ( ^Rj is independently a hydrocarbon group having 1 to 12 carbon atoms or a part of the hydrocarbon group is a nitrogen atom, oxygen. A group substituted with an atom or a sulfur atom) may be formed. ] Per fluorescent particles 1g, including the aggregated induced emissive molecule in an amount greater than 0.1 [mu] mol / g, the fluorescent particles of claim 1. The fluorescent particle according to any one of claims 1 to 2 , wherein the volume average particle diameter is 0.01 to 50 μm. The fluorescent particle according to any one of claims 1 to 3 , which is used for labeling. The swellable polymer particles, swollen with a liquid containing an aggregating induced luminescence molecules, viewed including the step of incorporating at least the aggregated induced luminescent molecules into swellable polymer particles,
The aggregation-induced luminescent molecule is at least one compound selected from the compounds represented by the following formulas (1), (2) and (3').
At least one of the swellable polymer particles selected from the group consisting of a polymerizable unsaturated aromatic compound, a polymerizable unsaturated carboxylic acid compound, a polymerizable unsaturated sulfonic acid compound or a salt thereof, and a polymerizable carboxylic acid ester compound. Particles obtained using seed compounds,
Method for producing fluorescent particles.

[In the formula (1), R ^{1 to} R ⁴ are independently hydrogen atoms, organic groups or organometallic groups, and a to d are independently integers of 1 to 5, and a plurality of Rs are used. ¹ , a plurality of R ² , a plurality of R ³ or a plurality of R ⁴ may be bonded to each other to form a ring, and R ¹ and R ² , R ² and R ⁴ , R ³ and R ⁴ , R ³ And R ¹ may be combined to form a ring. ]

[In formula (2), ^RA is independently a hydrogen atom or a hydrocarbon group having 1 to 12 carbon atoms, a is independently an integer of 1 to 5, and R ^B is independently containing an aromatic ring. It is an organic group and ^RC is an independent organic group. ]

[In formula (3'), ^Ra and R ^b are independently hydrogen atoms or organic groups having 1 to 20 carbon atoms, and R ^c and R ^d are independently hydrogen atoms, halogen atoms or carbon atoms, respectively. 20 is an organic group, ^{R e} is independently -COOR ^h or amino group ^{(R h} is a hydrogen atom or an organic group having 1 to 20 carbon atoms) is, ^{R f} is independently a hydrogen atom, It is an amide bond-containing group or an organic group having 1 to 20 carbon atoms which may have a halogen atom, an amino group and a protective group, and R ^g is independently a hydrogen atom, a halogen atom, a nitro group, a carboxyl group and an amino. It is a group or an organic group having 1 to 20 carbon atoms, m is an integer of 1 to 4, and R ^a and R ^c and / or R ^b and R ^d are bonded to each other and have a substituent R ^j . It may be a heterocycle having 5 or 6 constituent atoms containing at least one nitrogen atom ( ^Rj is independently a hydrocarbon group having 1 to 12 carbon atoms or a part of the hydrocarbon group is a nitrogen atom, oxygen. A group substituted with an atom or a sulfur atom) may be formed. ]