Polymer micelles containing nanoparticles in non-aqueous solution, methods for their preparation and use

Abstract

The present invention relates to a composition, comprising at least one micelle in non-aqueous solution, wherein the micelle encapsules one or more nanoparticles. Furthermore, the present invention relates to the use of such a composition and to methods for providing such a composition.

Claims

1. A method for providing a composition comprising at least one micelle encapsulating one or more nanoparticle(s) in non-aqueous solution, wherein in a first step the at least one micelle encapsulating the one or more nanoparticle(s) is formed in an aqueous solution where a surface of the at least one micelle encapsulating the one or more nanoparticle(s) is chemically-crosslinked; wherein in a second step the at least one micelle encapsulating the one or more nanoparticle(s) is transferred to a non-aqueous solution where the non-aqueous solution is at least partially miscible with the aqueous solution, has a higher boiling point than the aqueous solution, and does not form azeotropic phases with water; and wherein the second step further includes a removal of water to provide the composition comprising the at least one micelle encapsulating one or more nanoparticle(s) in the non-aqueous solution.

2. The method of claim 1, wherein the one or more nanoparticle(s) are luminescent.

3. The method of claim 2, wherein the one or more nanoparticle(s) comprise semi-conductor nanoparticle(s).

4. The method of claim 3, wherein the one or more nanoparticle(s) are doped by heavy-metal ions.

5. The method of claim 2, wherein the one or more nanoparticle(s) comprise a metal nanoparticle or a metal oxide nanoparticle.

6. The method of claim 1, wherein a material of the one or more nanoparticle(s) is defined by a formula M.sub.xN.sub.1-xA.sub.yB.sub.1-y, wherein M and N are independently selected from elements from group 8, 9, 10, 11, 12, 13, or 14 of the periodic table, wherein A and B are independently selected from elements from group 10, 11, 15, or 16 from the periodic table, and wherein x and y can be independently varied between 0 and 1.

7. The method of claim 6, wherein the one or more nanoparticle(s) comprise at least one shell.

8. The method of claim 7, wherein a material of the at least one shell is defined by a formula O.sub.1C.sub.zD.sub.1-z, wherein O is independently chosen from group 8, 9, 10, 11, 12, 13, or 14 from the periodic table, wherein C and D are independently chosen from group 10, 11, 15, or 16 from the periodic table; and wherein z independently assumes a value between 0 and 1 under a provision that M.sub.xN.sub.1-xA.sub.yB.sub.1-y and O.sub.1C.sub.zD.sub.1-z are not identical.

9. The method of claim 1, wherein the at least one micelle comprises a chemically crosslinked hydrophilic shell or a crosslinked hydrophilic shell obtained by polymerization.

10. The method of claim 1, wherein one or more micelle forming element(s) for the at least one micelle encapsulating one or more nanoparticle(s) include amphiphilic block co-polymers.

11. The method of claim 10, wherein the amphiphilic block co-polymers comprise at least two different polymer blocks of different compatibility towards solvents.

12. The method of claim 10, wherein the amphiphilic block co-polymers are one of linear, branched, star-shaped, or dendritic.

13. The method of claim 1, wherein the non-aqueous solution is selected from at least one of: dialkyl sulfoxides, e.g. dimethyl sulfoxide (DMSO); carboxylic acid amides, e.g. formamide, acetamide, N-methyl formamide, N-methyl acetamide, N,N-dimethyl formamide (DMF), N,N-dimethyl acetamide (DMA), or N-methyl pyrrolidone (NMP); phosphoric acid amides e.g. hexamethyl phosphoric acid triamide (HMPA); urea based solvents like 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU), 1,3-dimethyl-2-imidazolidinone (DMI), or tetramethyl urea (TEMUR); 1,4-dioxane; higher alkanols, e.g. 1-butanol; polyols, like ethylene glycol, 1,2-propanediol, 1,3-propanediol, glycerol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 1,2,4-butanetriol, diethylene glycol (DEG); ethers of polyols, e.g. 2-methoxyethanol, bis-(2-methoxyethyl) ether (diglyme); esters of polyols, e.g. diacetoxyethane; carboxylic acids, like formic acid, acetic acid, propanoic acid, lactic acid; primary, secondary and tertiary amines, diamines or triamines bearing branched, unbranched or cyclic alky groups on the nitrogen atoms and being liquids at ambient temperature; cyclic or bicylic amines, optionally bearing further heteroatoms, selected from N,O, or S in the ring, like piperidine, N-methyl piperidine, morpholine, N-methyl morpholine; annelated amidine bases, like 1,5-diazabicyclo(4.3.0)non-5-ene (DBN) or 1,8-diazabicyclo [5.4.0]undec-7-ene (DBU); optionally alkylated pyridines, e.g. pyridine, all isomers of picoline, lutidine and collidine; C1-2 nitro alkanes; ionic liquids, e.g. ammonium-based ionic liquids; imidazolium based ionic liquids like 1-allyl-3-methylimidazolium, 1-benzyl-3-methylimidazolium, 1-butyl-2,3-dimethylimidazolium, 1-butyl-3-methylimidazolium, 1-decyl-3-methylimidazolium, 1,3-didecyl-2-methylimidazolium, 1,3-dimethylimidazolium, 1,2-dimethyl-3-propylimidazolium, 1-dodecyl-3-methylimidazolium, 1-ethyl-2,3-dimethylimidazolium, 1-ethyl-3-methylimidazolium, 1-hexadecyl-3-methylimidazolium, 1-hexyl-3-methylimidazolium, 1-(2-hydroxyethyl)-3-methylimidazolium, 1-methyl-3-octadecylimidazolium, 1-methyl-3-octylimidazolium, 1-methyl-3-propylimidazolium salts, preferably with complex anions like e.g. tetrafluoroborate; piperidinium-based ionic liquids, like 1-butyl-1-methylpiperidinium, 1-methyl-1-propylpiperidinium; pyridinium-based ionic liquids, 1-alkyl-2-methylpyridinium, 1-alkyl-3-methylpyridinium, 1-alkyl-4-methylpyridinium, 1-butylpyridinium, 1-propylpyridinium, 1-ethylpyridinium, 1-hexylpyridinium salts, preferably with complex anions like e.g. tetrafluoroborate; pyrrolidinium-based ionic liquids, e.g. 1-butyl-1-methylpyrrolidinium, 1-ethyl-1-methylpyrrolidinium, 1-methyl-1-propylpyrrolidinium salts, preferably with complex anions, which in combination with the cation renders the ionic liquid water miscible, like e.g. tetrafluoroborate, or mixtures of said solvents.

14. The method of claim 1, further comprising conjugating the composition comprising the at least one micelle encapsulating one or more nanoparticle(s) in the non-aqueous solution to a biomolecule of a plurality of biomolecules.

15. The method of claim 1, wherein the removal of water is via distillation at or below atmospheric pressure.

16. The method of claim 1, wherein the removal of water is via one or more of azeotropic removal of water, diffusion or permeation across porous or non-porous membranes, and chemical drying.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows DLS-Data for micellar encapsulated nanoparticles in methanol and water.

(2) FIG. 2 shows the absorption spectrum (left) and emission spectrum (right) of native CdSeTe/CdS nanoparticles in chloroform and of micellar encpasulated CdSeTe/CdS nanoparticles in methanol.

(3) FIG. 3 shows the absorption spectrum (left) and emission spectrum (right) of native CdSe/CdS/ZnS nanoparticles in chloroform and of micellar encpasulated CdSe/CdS/ZnS nanoparticles in methanol.

(4) FIG. 4 shows agarose gel analysis of fluorescent quantum dot-rod nanoparticle-DNA conjugates. Conjugation was performed in aqueous solution (left), in 1,2-propanediol (middle) and 1,3-butanediol (right). Success of DNA binding is evident from increased movement from top (cathode) to the bottom (anode) of the gel.

(5) FIG. 5 shows spot tests from functional analysis of fluorescent quantum dot-rod nanoparticle-DNA conjugates. Conjugation was performed in aqueous solution (left), in 1,2-propanediol (middle) and 1,3-butanediol (right). Success of DNA binding is evident from accumulation of fluorescent quantum dot-rod nanoparticle-DNA conjugates on spotted complementary DNA.

(6) FIG. 6 shows the dynamic light scattering of micelle encapsulated trioctylamine coated CdSeTe/CdS nanoparticles in methanol.

(7) FIG. 7 shows the absorption spectra of TOA coated CdSeTe/CdS nanoparticles in chloroform (solid line) and the micelle encapsulated TOA coated CdSeTe/CdS nanoparticles in methanol (dotted line).

(8) FIG. 8 shows the fluorescence spectra of TOA coated CdSeTe/CdS nanoparticles in chloroform (solid line) and the micelle encapsulated TOA coated CdSeTe/CdS nanoparticles in methanol (dotted line).

(9) FIG. 9 shows the dynamic light scattering of micelle encapsulated PI-DETA coated

(10) CdSeTe/CdS nanoparticles in methanol

(11) FIG. 10 shows the absorption spectra of PI-DETA coated CdSeTe/CdS nanoparticles in chloroform (solid line) and the micelle encapsulated PI-DETA coated CdSeTe/CdS nanoparticles in methanol (dotted line).

(12) FIG. 11 shows the fluorescence spectra of PI-DETA coated CdSeTe/CdS nanoparticles in chloroform (solid line) and the micelle encapsulated PI-DETA coated CdSeTe/CdS nanoparticles in methanol (dotted line).

(13) FIG. 12 shows the dynamic light scattering of micelle encapsulated PI-DETA coated CdSe/CdS/ZnS nanoparticles in methanol.

(14) FIG. 13 shows the absorption spectra of PI-DETA coated CdSe/CdS/ZnS nanoparticles in chloroform (solid line) and the micelle encapsulated PI-DETA coated CdSe/CdS/ZnS nanoparticles in methanol (dotted line).

(15) FIG. 14 shows the fluorescence spectra of PI-DETA coated CdSe/CdS/ZnS nanoparticles in chloroform (solid line) and the micelle encapsulated PI-DETA coated CdSe/CdS/ZnS nanoparticles in methanol (dotted line).

(16) FIG. 15 shows the dynamic light scattering of micelle encapsulated CdSe/CdS nanoparticles in methanol.

(17) FIG. 16 shows the absorption spectra of CdSe/CdS nanoparticles in chloroform (solid line) and the micelle encapsulated CdSe/CdS nanoparticles in methanol (dotted line).

(18) FIG. 17 shows the fluorescence spectra of native CdSe/CdS nanoparticles (solid line) and the micelle encapsulated CdSe/CdS nanoparticles in methanol (dotted line).

(19) FIG. 18 shows the dynamic light scattering of micelle encapsulated CdSe/CdS rod-shaped nanoparticles in methanol.

(20) FIG. 19 shows the absorption spectra of CdSe/CdS rod-shaped nanoparticles in chloroform (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in methanol (dotted line).

(21) FIG. 20 shows the fluorescence spectra of native CdSe/CdS rod-shaped nanoparticles (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in methanol (dotted line).

(22) FIG. 21 shows the dynamic light scattering of micelle encapsulated CdSe/CdS rod-shaped nanoparticles in acetonitrile.

(23) FIG. 22 shows the absorption spectra of CdSe/CdS rod-shaped nanoparticles in n-hexane (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in acetonitrile (dotted line).

(24) FIG. 23 shows the fluorescence spectra of native CdSe/CdS rod-shaped nanoparticles (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in acetonitrile (dotted line).

(25) FIG. 24 shows the dynamic light scattering of micelle encapsulated CdSe/CdS rod-shaped nanoparticles in dimethyl sulfoxide.

(26) FIG. 25 shows the absoprtion spectra of CdSe/CdS rod-shaped nanoparticles in n-hexane (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in dimethyl sulfoxide (dotted line).

(27) FIG. 26 shows the fluorescence spectra of native CdSe/CdS rod-shaped nanoparticles (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in dimethyl sulfoxide (dotted line).

(28) FIG. 27 shows the dynamic light scattering of micelle encapsulated CdSe/CdS rod-shaped nanoparticles in N,N-dimethylformamide.

(29) FIG. 28 shows the absoprtion spectra of CdSe/CdS rod-shaped nanoparticles in n-hexane (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in N,N-dimethylformamide (dotted line).

(30) FIG. 29 shows the fluorescence spectra of native CdSe/CdS rod-shaped nanoparticles (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in N,N-dimethylformamide (dotted line).

(31) FIG. 30 shows the dynamic light scattering of micelle encapsulated CdSe/CdS rod-shaped nanoparticles in the ionic liquid [BMIM][BF.sub.4].

(32) FIG. 31 shows the absoprtion spectra of CdSe/CdS rod-shaped nanoparticles in n-hexane (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in [BMIM][BF4] (dotted line).

(33) FIG. 32 shows the fluorescence spectra of native CdSe/CdS rod-shaped nanoparticles (solid line) and the micelle encapsulated CdSe/CdS rod-shaped nanoparticles in [BMIM][BF.sub.4] (dotted line).

(34) FIG. 33 shows the dynamic light scattering of micelle encapsulated InP/ZnS nanoparticles in methanol.

(35) FIG. 34 shows the absorption spectra of InP/ZnS nanoparticles in toluene (solid line) and the micelle encapsulated InP/ZnS nanoparticles in methanol (dotted line).

(36) FIG. 35 shows the fluorescence spectra of native InP/ZnS nanoparticles (solid line) and the micelle encapsulated InP/ZnS nanoparticles in methanol (dotted line).

(37) FIG. 36 shows the absorption spectra of native InP/ZnS nanoparticles (solid line), hexadecylamine coated InP/ZnS nanoparticles (dashed line) and the micelle encapsulated hexadecylamine coated InP/ZnS nanoparticles in methanol (dotted line).

(38) FIG. 37 shows the fluorescence spectra of native InP/ZnS nanoparticles (solid line), hexadecylamine coated InP/ZnS nanoparticles (dashed line) and the micelle encapsulated hexadecylamine coated InP/ZnS nanoparticles in methanol (dotted line).

(39) FIG. 38 shows the dynamic light scattering of micelle encapsulated CulnSe/ZnS nanoparticles in methanol.

(40) FIG. 39 shows the absorption spectra of CuInSe/ZnS nanoparticles in chloroform (solid line) and the micelle encapsulated CulnSe/ZnS nanoparticles in methanol (dotted line).

(41) FIG. 40 shows the fluorescence spectra of native CuInSe/ZnS nanoparticles (solid line) and the micelle encapsulated CulnSe/ZnS nanoparticles in methanol (dotted line).

(42) FIG. 41 shows the dynamic light scattering of micelle encapsulated Au nanoparticles in methanol.

(43) FIG. 42 shows the absorption spectra of Au nanoparticles in toluene (solid line) and the micelle encapsulated Au nanoparticles in methanol (dotted line).

(44) FIG. 43 shows the dynamic light scattering of micelle encapsulated Au nanoparticles in acetonitrile.

(45) FIG. 44 shows the absorption spectra of Au nanoparticles in toluene (solid line) and the micelle encapsulated Au nanoparticles in acetonitrile (dotted line).

(46) FIG. 45 shows the dynamic light scattering of micelle encapsulated Au nanoparticles in N,N-dimethylformamide.

(47) FIG. 46 shows the absorption spectra of Au nanoparticles in toluene (solid line) and the micelle encapsulated Au nanoparticles in N,N-dimethylformamide (dotted line).

(48) FIG. 47 shows the dynamic light scattering of micelle encapsulated Au nanoparticles in DMSO.

(49) FIG. 48 shows the absorption spectra of Au nanoparticles in toluene (solid line) and the micelle encapsulated Au nanoparticles in DMSO (dotted line).

(50) FIG. 49 shows the dynamic light scattering of micelle encapsulated Au nanoparticles in [BMIM][BF.sub.4].

(51) FIG. 50 shows the absorption spectra of Au nanoparticles in toluene (solid line) and the encapsulated Au nanoparticles in ionic liquid [BMIM][BF.sub.4] (dotted line).

(52) FIG. 51 shows the dynamic light scattering of micelle encapsulated Fe.sub.xO.sub.y nanoparticles in methanol.

(53) FIG. 52 shows the dynamic light scattering of micelle encapsulated Fe.sub.xO.sub.y nanoparticles in DMSO.

(54) FIG. 53 shows the dynamic light scattering of micelle encapsulated Fe.sub.xO.sub.y nanoparticles in N,N-dimethylformamide.

(55) FIG. 54 shows the dynamic light scattering of micelle encapsulated Fe.sub.xO.sub.y nanoparticles in [BMIM] [BF.sub.4].

(56) FIG. 55 shows the dynamic light scattering of micelle encapsulated NaYF.sub.4:Yb/NaYF.sub.4 nanoparticles in methanol.

(57) FIG. 56 shows the fluorescence spectra of native NaYF.sub.4:Yb/NaYF.sub.4 nanoparticles (solid line) and the micelle encapsulated NaYF.sub.4:Yb/NaYF.sub.4 nanoparticles in methanol (dotted line); Excitation at 978 nm.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

(58) Definitions

(59) In particular, the present invention relates to nanoparticles, possessing superparamagnetic, plasmonic, or fluorescent properties, including semiconductor nanoparticles, metallic nanoparticles, oxidic nanoparticles, or rare earth doped nanoparticles encapsulated in micelles in non-aqueous solutions.

(60) Nanoparticles within the scope of the invention are spatial objects, which have dimensions of 1 to 1000 nm in at least one dimension. Said nanoparticles may occur as solid particles, as well as hollow structures in various shapes, which include, but are not restricted to, spherical, elongated, i.e. rod-like, elliptical, egg-like, dumbbell-shaped, circular or torus-like, cubic or cuboid, prismatic or cylindrical shapes, regularly or irregularly multi-faceted geometrical bodies, polygonal plates of e.g. triangular, square, or hexagonal shapes, leaflets, multi-armed or star-shaped bodies, e.g. tetrapods, or wire-like shapes, and the like. Said nanoparticles may be optionally shielded with at least one shell comprised of a different material than the core.

(61) The term nanoparticle is inter alia also covering the term nano-crystal or ultra-small particle.

(62) Before the present invention is described in detail, it is to be understood that this invention is not limited to the particular methodology, devices, or compositions described, as such methods, devices, or compositions can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.

(63) Use of the singular forms a, an, and the include plural references unless the context clearly dictates otherwise. Thus, for example, reference to a cell includes a plurality of cells, reference to a nanoparticle includes a plurality of such nanoparticles, reference to an biomolecule like i.e. DNA, protein, peptide, carbohydrate or the like includes the plurality of biomolecules, and the like.

(64) Unless defined otherwise or the context clearly dictates otherwise, 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 invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the invention, the preferred methods and materials are now described.

(65) All publications mentioned herein are hereby incorporated by reference for the purpose of disclosing and describing the particular materials and methodologies for which the reference was cited. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

(66) In describing the present invention, the following terms will be employed, and are intended to be defined as indicated below.

(67) The term nanoparticle refers to a particle, generally a semiconductor particle, or a nanocrystalline non-semiconductor-based particle, in particular, a lanthanide-doped metal oxide particle or lanthanide-doped salt-like particle, or an oxidic or a metallic particle, having a diameter in the range of about 1 nm to about 1000 nm, preferably in the range of about 2 nm to about 50 nm, more preferably in the range of about 2 nm to about 20 nm (for example about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nm).

(68) The terms semiconductor nanocrystal and quantum dot, quantum rod, quantum dot rod or the acronyms QD, QDs, QR, QRs,QDR, QDRs, SCNC or SCNCs are used interchangeably herein to refer to semiconductor nanoparticles composed of an inorganic crystalline material that is luminescent (i.e., they are capable of emitting 'electromagnetic radiation upon excitation), and include an inner core of one or more first semiconductor materials that is optionally contained within an overcoating or shell of a second semiconductor material. A semiconductor nanocrystal core surrounded by a semiconductor shell is referred to as a core/shell semiconductor nanocrystal.

(69) Semiconductors are especially defined by their electrical conductivity, which lies between the one of a conductor and the one of an insulator. Semiconductors can be luminescent, but this is not mandatory

(70) For example, the surrounding shell material may have a bandgap energy that is larger than the bandgap energy of the core material and may be chosen to have an atomic spacing close to that of the core substrate.

(71) Suitable semiconductor materials for the core and/or shell may include, but not limited to, the following materials comprised of a first element selected from Groups 2 or 12 of the Periodic Table of the Elements and a second element selected from Group 16 (e.g., ZnS, ZnSe, ZnTe, CdS, CdSe, CdTe, HgS, HgSe, HgTe, MgS, MgSe, MgTe, CaS, CaSe, CaTe, SrS, SrSe, SrTe, BaS, BaSe, BaTe, and the like as well as mixtures thereof); materials comprised of a first element selected from Group 13 of the Periodic Table of the Elements and a second element selected from Group 15 (GaN, GaP, GaAs, GaSb, InN, InP, InAs, InSb, and the like as well as mixtures thereof); materials comprised of a Group 14 element (Ge, Si, and the like as well as mixtures thereof); materials such as PbS, PbSe, PbTe and the like as well as mixtures thereof, materials such as CuInS, CuInGaS and the like as well as mixtures thereof; and alloys and mixtures thereof. As used herein, all reference to the Periodic Table of the Elements and groups thereof is to the new IUPAC system for numbering element groups, as set forth in the Handbook of Chemistry and Physics, 81st Edition (CRC Press, 2000).

(72) An SCNC is optionally surrounded by a coat of an organic capping agent. The organic capping agent may be any number of materials, but has an affinity for the SCNC surface. In general, the capping agent can be an isolated organic molecule, a polymer (or a monomer for a polymerization reaction), an inorganic complex, or an extended crystalline structure. The coat can be used to convey solubility, e.g., the ability to disperse a coated SCNC homogeneously into a chosen solvent, functionality, binding properties, or the like. In addition, the coat can be used to tailor the optical properties of the SCNC.

(73) Thus, the terms semiconductor nanocrystal, SCNC, and quantum dot as used herein include a coated SCNC core, as well as a core/shell SCNC.

(74) Monodisperse particles include a population of particles wherein at least about 60% of the particles in the population, more preferably about 75 to about 90, or any integer therebetween, percent of the particles in the population fall within a specified particle size range. A population of monodisperse particles deviates less than 10% rms (root-mean-square) in diameter, and preferably deviates less than 5% rms.

(75) The phrase one or more sizes of SCNCs is used synonymously with the phrase one or more particle size distributions of SCNCs. One of ordinary skill in the art will realize that particular sizes of SCNCs are actually obtained as particle size distributions.

(76) By luminescence is meant the process of emitting electromagnetic radiation (light) from an object. Luminescence results when a system undergoes a transition from an excited state to a lower energy state with a corresponding release of energy in the form of a photon. These energy states can be electronic, vibrational, rotational, or any combination thereof. The transition responsible for luminescence can be stimulated through the release of energy stored in the system chemically or added to the system from an external source. The external source of energy can be of a variety of types including chemical, thermal, electrical, magnetic, electromagnetic, and physical, or any other type of energy source capable of causing a system to be excited into a state higher in energy than the ground state. For example, a system can be excited by absorbing at least one photon of light, by being placed in an electrical field, or through a chemical oxidation-reduction reaction. The energy of the photons emitted during luminescence can be in a range from low-energy microwave radiation to high-energy x-ray radiation. Typically, luminescence refers to photons in the range from UV to IR radiation.

(77) Biomolecule refers to any kind of molecule used in biological applications, including but not limited to, optionally glycosylated proteins, e.g. antibodies, nanobodies, antibody fragments, like single-chain antibodies, fab fragments, viral proteins, lectins, peptides, including polyaminoacids, nucleic acids, including desoxyribonucleic acids (DNA), ribonucleic acids (RNA, siRNA, miRNA), locked nucleic acid (LNA), aptamers, lipids, steroids, messenger substances, prions, carbohydrates, small molecules and the like.

(78) Complementary or substantially complementary refers to the ability to hybridize or base pair between nucleotides or nucleic acids, such as, for instance, between the two strands of a double stranded DNA molecule or between a polynucleotide primer and a primer binding site on a single stranded nucleic acid to be sequenced or amplified. Complementary nucleotides are, generally, Adenine [A] and Thymine [T], or A and Uracil [U], or Cytosine [C] and Guanine [G]. Two single-stranded RNA or DNA molecules are said to be substantially complementary when the nucleotides of one strand, optimally aligned and compared and with appropriate nucleotide insertions or deletions, pair with at least about 80% of the nucleotides of the other strand, usually at least about 90% to 95%, and more preferably from about 98 to 100%.

(79) The term aptamer (or nucleic acid antibody) is used herein to refer to a single- or double-stranded polynucleotide that recognizes and binds to a desired target molecule by virtue of its shape (c.f. e.g., PCT Publication Nos. WO 92/14843, WO 91/19813, and WO 92/05285).

(80) According to the invention, the problems described above are solved by the encapsulation of nanoparticles into micelles in non-aqueous solutions.

(81) The term encapsulation refers to micellization as well as to cross-linking a previously formed micelle or micelles.

(82) The term micelle may be generally understood as an aggregate of colloidal dimensions of surfactant molecules dispersed in a liquid, and the term surfactant as a surface-active substance, lowering the surface tension of liquid, usually organic compounds that are amphiphilic.

(83) The term transfection may be generally understood as the deliberate or also undeliberate introduction of material like the micelle-encapsulated nanoparticles into cells.

(84) The term labeling may be generally understood as the deliberate or also underliberate attaching of material like the micelle-encapsulated nanoparticles to a cell or a biological molecule or a biological sample. The material may be e.g. attached onto the cell or into the cell.

(85) Transfection and labeling of a cell or a biological molecule or a biological sample are both examples for a biological application.

(86) The term biological event can be defined as including an interaction of biological moieties, a biological process, an alteration in the structures of a biological compound or, an alteration in a biological process.

(87) The term biological sample can be defined as a sample of tissue or fluid isolated from an individual, including but not limited to, for example, plasma, serum, spinal fluid, semen, lymph fluid, the external sections of the skin, respiratory, intestinal, and genitourinary tracts, tears, saliva, milk, blood cells, tumors, organs, and also samples of in vitro cell culture constituents (including but not limited to conditioned medium resulting from the growth of cells in cell culture medium, putatively virally infected cells, recombinant cells, and cell components).

(88) In particular, by the present invention a composition is provided, comprising at least one micelle in non-aqueous solution, wherein the micelle encapsulates one or more nanoparticle(s).

(89) The nanoparticles can be monodisperse nanoparticles.

(90) Synthesis of nanoparticles in reverse micelles have been described (F. T. Quinlan, J. Kuther, W. Tremel, W. Knoll, S. Risbud, P. Stroeve; Reverse Micelle Synthesis and Characterization of ZnSe Nanoparticles, Langmuir 2000, 16, 4049-4051.) (M.-L. Wu, D.-H. Chen, T.-C. Huang; Synthesis of Au/Pd Bimetallic Nanoparticles in Reverse Micelles, Langmuir 2001, 17, 3877-3883.) (Dongbai Zhang, Limin Qi, Jiming Ma, Humin Cheng; Formation of crystalline nanosized titania in reverse micelles at room temperature, J. Mater. Chem., 2002, 12, 3677-3680), (Christopher E. Bunker et al. Formation of Cadmium Sulfide Nanoparticles in Reverse Micelles: Extreme Sensitivity to Preparation Procedure, Langmuir 2004, 20, 5642-5644), however the formation of micelles around freely dispersed pre-formed nanoparticles in non-aqueous solvents is novel. Surprisingly, the micellar compositions of said nanoparticles thus obtained, maintain their integrity and properties when transferred into different non-aqueous and aqueous solvents. Furthermore it was found that polymer micelles containing nanoparticles in an aqueous preparation can be transferred into non-aqueous solutions by removing the water, virtually without impairing their properties.

(91) Within the scope of the invention, nanoparticles suitable for the encapsulation in micelles in non-aqueous solutions may bear a hydrophobic surface originating from said growth process.

(92) Said nanoparticles as derived from their growth solution may occur as solid particles, as well as hollow structures in various shapes, which include, but are not restricted to, spherical, rod-like, elliptical, egg-like, circular or torus-like, cubic or cuboid, or cylindrical shapes, regularly or irregularly multi-faceted geometrical bodies, polygonal plates of e.g. triangular, square, or hexagonal shapes, multi-armed or star-shaped bodies, e.g. tetrapods, wire-like shapes, and the like.

(93) Said nanoparticles may exhibit characteristic physico-chemical attributes on interacting with their environment, e.g. absorbance of parts of the electromagnetic spectrum and fluorescence emission on excitation with parts of the electromagnetic spectrum, magnetism, paramagnetism, superparamagnetism, plasmon resonance, Raman-Effect enhancement or combinations of said properties.

(94) Nanoparticles within the scope of the invention may include, but are not limited to semiconductor quantum dots or quantum rods, or combinations thereof; salt-like lattices, e.g. phosphates, vanadates, wolframates, fluorides, oxychlorides and the like; metals, including gold and silver, or metal oxides, e.g. iron oxides in various modifications.

(95) Said semiconductor nanoparticles may be e.g. described by the formula M.sub.xN.sub.1-xA.sub.yB.sub.1-y, wherein M and N are independently selected from elements from group 8, 9, 10, 11, 12, 13 or 14 of the periodic table, e.g. but not limited to Fe, Co, Ni, Pt, Cu, Zn, Cd, Al, Ga, In, Ge, Sn or Pb, and A and B are independently selected from elements from group 10, 11, 15 or 16 from the periodic table, e.g. but not limited to Pd, Pt, N, P, As, Sb, O, S, Se or Te, and wherein x and y can be independently varied between 0 and 1.

(96) Said semiconductor nanoparticles may be optionally shielded by at least one shell comprising the formula O.sub.1C.sub.zD.sub.1-z, wherein O can be independently chosen from group 8, 9, 10, 11, 12, 13 or 14 from the periodic table, e.g. but not limited to Fe, Co, Ni, Pt, Cu, Zn, Cd, Al, Ga, In, Ge, Sn or Pb, and C and D can be independently chosen from group 10, 11, 15 or 16 from the periodic table, e.g. but not limited to Pd, Pt, N, P, As, Sb, O, S, Se or Te, and wherein z may independently assume a value between 0 and 1 under the proviso that the compositions of M.sub.xN.sub.1-xA.sub.yB.sub.1-y and O.sub.1C.sub.zD.sub.1-z are not identical.

(97) Each of the structural components, i.e. core or shells can be optionally doped by heavy-metal ions like Ag, Cu, Co, or Mn, thus comprising at least one said heavy-metal atom per nanoparticle.

(98) Typical compositions may include but are not limited to ZnS, ZnSe, ZnTe, CdS, CdSe, CdTe, HgS, HgSe, HgTe, MgS, MgSe, MgTe, CaS, CaSe, CaTe, SrS, SrTe, BaS, BaSe, BaTe, GaN, GaP, GaAs, GaSb, InN, InP, InAs, InSb, Ge, Si, PbS, PbSe, PbTe, CuInS, CuInGaS and an alloy or mixture thereof.

(99) Said shells may independently emulate the shape of the core particle, or may be of different shapes. Exemplarily, if the core is spherical, elongated or tripod-, tetrapod-, hexapod-, octapod- or star-shaped, a core/shell particle may be comprised of core(spherical)/shell(spherical), core(elongated)/shell(elongated), core(spherical)/shell(elongated), core(spherical)/shell(tetrapod). Exemplarily, a core/shell/shell nanoparticle may be comprised of core(spherical)/shell(spherical)/shell(spherical), core(spherical)/shell(spherical)/shell(elongated), core(spherical)/shell(spherical)/shell(tetrapod), core(spherical)/shell(elongated)/shell(elongated), core(elongated)/shell(elongated)/shell(elongated). Exemplarily, a core/shell/shell/shell nanoparticle may be comprised of core(spherical)/shell(spherical)/shell(spherical)/shell(spherical), core(spherical)/shell(spherical)/shell(spherical)/shell(elongated), core(spherical)/shell(spherical)/shell(elongated)/shell(elongated), core(spherical)/shell(elongated)/shell(elongated)/shell(elongated), core(elongated)/shell(elongated)/shell(elongated)/shell(elongated).

(100) Said rare earth doped nanoparticles may exhibit optionally luminescent and/or paramagnetic properties.

(101) Said luminescent rare earth doped nanoparticles are particles wherein a core is comprised of a first metal salt or oxide, whichafter excitationis suited to transfer the excitation energy to a second luminescent metal salt, or to a plurality of luminescent metal salts, which emits said excitation energy as luminescence.

(102) Luminescent nanoparticles are comprised of a core and optionally shielded by at least one shell, wherein the core metal and the shell metal are selected from the elements Ce (58), Pr (59), Nd (60), Sm (62), Eu (63), Gd (64), Tb (65), Dy (66), Ho (67), Er (68), Tm (69) or Yb (70). Specific examples of luminescent shell materials are for instance LiI:Eu; Nal:TI; Csl:TI; Csl:Na; LiF:Mg; LiF:Mg,Ti; LiF:Mg,Na; KMgF.sub.3: Mn; BaFCl:Eu; BaFCl:Sm; BaFBr:Eu; BaFCl.sub.0.5Br.sub.0.5:Sm; BaY.sub.2F.sub.8:A (A=Pr, Tm, Er, Ce); BaSi.sub.2O.sub.5:Pb; BaMg.sub.2Al.sub.16O.sub.27:Eu; BaMgAl.sub.14O.sub.23: Eu; BaMgAl.sub.10O.sub.17:Eu; BaMgAl.sub.2O.sub.3:Eu; Ba.sub.2P.sub.2O.sub.7:Ti; (Ba,Zn,Mg).sub.3Si.sub.2O.sub.7:Pb; Ce(Mg,Ba)Al.sub.11O.sub.19; Ce.sub.0.65Tb.sub.0.35MgAl.sub.11O.sub.19:Ce,Tb; MgAl.sub.11O.sub.19Ce,Tb; MgF.sub.2:Mn; MgS:Eu; MgS:Ce; MgS:Sm; MgS:(Sm,Ce); (Mg,Ca)S:Eu; MgSiO.sub.3:Mn; 3.5MgO.0.5MgF.sub.2.GeO.sub.2:Mn; MgWO.sub.4:Sm; MgWO.sub.4:Pb; 6MgO.As.sub.2O.sub.5:Mn; (Zn,Mg) F.sub.2:Mn; (Zn.sub.4Be)SO.sub.4:Mn; Zn.sub.2SiO.sub.4:Mn; Zn.sub.2SiO.sub.4:Mn,As; Zn.sub.3(PO.sub.4).sub.2:Mn; CdBO.sub.4:Mn; CaF.sub.2:Mn; CaF.sub.2:Dy; CaS:A (A=Lanthanide,Bi); (Ca,Sr)S:Bi; CaWO.sub.4:Pb; CaWO.sub.4:Sm; CaSO.sub.4:A (A=Mn, lanthanide); 3Ca.sub.3(PO.sub.4).sub.2.Ca(F,Cl).sub.2:Sb,Mn; CaSiO.sub.3:Mn,Pb; Ca.sub.2Al.sub.2Si.sub.2O.sub.7:Ce; (Ca,Mg)SiO.sub.3:Ce; (Ca,Mg)SiO.sub.3:Ti; 2SrO.6(B.sub.2O.sub.3).SrF.sub.2Eu; 3Sr.sub.3(PO.sub.4).sub.2.CaCl.sub.2:Eu; A.sub.3(PO.sub.4).sub.2.ACl.sub.2:Eu (A=Sr, Ca, Ba); (Sr,Mg).sub.2P.sub.2O.sub.7:Eu; (Sr,Mg).sub.3(PO.sub.4).sub.2:Sn; SrS:Ce; SrS:Sm,Ce; SrS:Sm; SrS:Eu; SrS:Eu,Sm; SrS:Cu,Ag; Sr.sub.2P.sub.2O.sub.7:Sn; Sr.sub.2P.sub.2O.sub.7:Eu; Sr.sub.4Al.sub.14O.sub.25:Eu; SrGa.sub.2S.sub.4:A (A=lanthanide, Pb); SrGa.sub.2S.sub.4:Pb; Sr.sub.3Gd.sub.2Si.sub.6O.sub.18:Pb, Mn; YF.sub.3:Yb, Er; YF.sub.3:Ln (Ln=lanthanide); YLiF.sub.4:Ln (Ln=lanthanide); Y.sub.3Al.sub.5O.sub.12:Ln (Ln=lanthanide); YAl.sub.13(BO.sub.4).sub.3:Nd, Yb; (Y,Ga)BO.sub.3:Eu; (Y,Gd)BO.sub.3:Eu; Y.sub.2Al.sub.3Ga.sub.2O.sub.12:Tb; Y.sub.2SiO.sub.5:Ln (Ln=lanthanide); Y.sub.2O.sub.2S:Ln (Ln=lanthanide); YVO.sub.4:A (A=lanthanide, In); Y(P,V)O.sub.4:Eu; YTaO.sub.4:Nb; YAlO.sub.3:A (A=Pr, Tm, Er, Ce); YOCl:Yb,Er; Ln(PO.sub.4:Ce,Tb (Ln=lanthanide or mixture of lanthanides); LuVO.sub.4:Eu; GdVO.sub.4:Eu; Gd.sub.2O.sub.2S:Tb; GdMgB.sub.5O.sub.10:Ce,Tb; LaOBr:Tb; La.sub.2O.sub.2S:Tb; LaF.sub.3:Nd,Ce;BaYb.sub.2F.sub.8:Eu; NaYF.sub.4:Yb,Er; NaYF.sub.4:Yb,Tm, NaGdF.sub.4:Yb,Er; NaGdF.sub.4:Yb,Tm, NaLaF.sub.4:Yb,Er; NaLaF.sub.4:Yb,Tm, LaF.sub.3:Yb,Er,Tm; BaYF.sub.3:Yb,Er; GaN:A (A=Pr, Eu, Er, Tm); Bi.sub.4Ge.sub.3O.sub.12; LiNbO.sub.3:Nd,Yb; LiNbO.sub.3:Er; LiCaAlF.sub.6:Ce; LiSrAlF.sub.6:Ce; LiLuF.sub.4:A (A=Pr, Tm, Er, Ce); Li.sub.2B.sub.4O.sub.7:Mn, Y.sub.2O.sub.2S:Eu, Y.sub.2SiO.sub.5:Eu, CaSiO.sub.3:Ln, CaS:Ln, wherein Ln=one, two or more lanthanides as well as mixtures thereof.

(103) If classified according to the host lattice type the following preferred embodiments can also be enumerated. 1. Halides: for instance XY.sub.2 (XMg, Ca, Sr, Ba; YF, Cl, I), CaF:Eu (II), BaF:Eu; BaMgF.sub.4:Eu; LiBaF.sub.3:Eu; SrF:Eu; SrBaF.sub.2Eu; CaBr.sub.2:EuSiO.sub.2; CaCl.sub.2:Eu; CaCl.sub.2:EuSiO.sub.2; CaCl.sub.2:Eu,MnSiO.sub.2; CaI.sub.2:Eu; CaI.sub.2:Eu, Mn; KMgF.sub.3:Eu; SrF.sub.2:Eu (II), BaF.sub.2:Eu (II), YF.sub.3, NaYF.sub.4; MgF.sub.2:Mn; MgF.sub.2:Ln (Ln=lanthanide (s)) as well as mixtures thereof. 2. Earth alkaline sulfates: for instance XSO.sub.4 (XMg, Ca, Sr, Ba), SrSO.sub.4:Eu, SrSO.sub.4:Eu,Mn, BaSO.sub.4:Eu, BaSO.sub.4:Eu,Mn, CaSO.sub.4, CaSO.sub.4:Eu, CaSO.sub.4:Eu,Mn, as well as mixed earth alkaline sulfates, also in combination with magnesium, e.g. Ca,MgSO.sub.4:Eu,Mn as well as mixtures thereof. 3. Phosphates and halophosphates: for instance CaPO.sub.4:Ce,Mn, Ca.sub.5(PO.sub.4).sub.3Cl:Ce,Mn, Ca.sub.5(PO.sub.4).sub.3F:Ce,Mn, Sr(PO.sub.4:Ce,Mn, Sr.sub.5(PO.sub.4).sub.3Cl:Ce,Mn, Sr.sub.5(PO.sub.4).sub.3F:Ce,Mn, the latter also codoped with Eu (II) or codoped with EU,Mn, -Ca.sub.3(PO.sub.4).sub.2: Eu; -Ca.sub.3(PO.sub.4).sub.2:Eu,Mn; Ca.sub.5(PO.sub.4).sub.3Cl:Eu; Sr.sub.5(PO.sub.4).sub.3Cl:Eu; Ba.sub.10(PO.sub.4).sub.6Cl:Eu; Ba.sub.10(PO.sub.4).sub.6Cl:Eu,Mn, Ca.sub.2Ba.sub.3(PO.sub.4).sub.3Cl:Eu; Ca.sub.5(PO.sub.4).sub.3F:Eu.sup.2+X.sup.3+; Sr.sub.5(PO.sub.4).sub.3F:Eu.sup.2+X.sup.3+(XNd, Er, Ho, Tb); Ba.sub.5(PO.sub.4).sub.3Cl:Eu; -Ca.sub.3(PO.sub.4).sub.2:Eu; CaB.sub.2P.sub.2O.sub.9:Eu; Ca.sub.2P.sub.2O.sub.7:Eu; Ca.sub.2P.sub.2O.sub.7:Eu, Mn; Sr.sub.10(PO.sub.4).sub.6Cl:Eu; (Sr, Ca, Ba, Mg).sub.10(PO.sub.4).sub.6Cl:Eu; La(PO.sub.4:Ce; CePO.sub.4; LaPO.sub.4:Eu, LaPO.sub.4:Ce, LaPO.sub.4:Ce,Tb, CePO.sub.4:Tb as well as mixtures thereof. 4. Borates: for instance LaBO.sub.3; LaBO.sub.3:Ce; ScBO.sub.3:Ce; YAlBO.sub.3:Ce; YBO.sub.3:Ce; Ca.sub.2B.sub.5O.sub.9Cl:Eu; xEuO.yNa.sub.2O.zB.sub.2O.sub.3as well as mixtures thereof. 5. Vanadates: for instance YVO.sub.4, YVO.sub.4:Eu, YVO.sub.4:Dy, YVO.sub.4:Sm; YVO.sub.4:Bi; YVO.sub.4:Bi,Eu, YVO.sub.4:Bi,Dy, YVO.sub.4:Bi,Sm; YVO.sub.4:Tm, YVO.sub.4:Bi,Tm GdVO.sub.4, GdVO.sub.4:Eu, GdVO.sub.4:Dy, GdVO.sub.4:Sm GdVO.sub.4:Bi; GdVO.sub.4:Bi,Eu, GdVO.sub.4:Bi,Dy, GdVO.sub.4:Bi,Sm; Gd(PV)O.sub.4, Gd(PV)O.sub.4:Eu, Gd(PV)O.sub.4:Dy, Gd(PV)O.sub.4:Sm Gd(PV)O.sub.4:Bi; Gd(PV)O.sub.4:Bi,Eu, Gd(PV)O.sub.4:Bi,Dy, Gd(PV)O.sub.4:Bi,Sm as well as mixtures thereof. 6. Aluminates: for instance MgAl.sub.2O.sub.4:Eu; CaAl.sub.2O.sub.4:Eu; SrAl.sub.2O.sub.4:Eu; BaAl.sub.2O.sub.4:Eu; LaMgAl.sub.11O.sub.19:Eu; BaMgAl.sub.10O.sub.17:Eu; BaMgAl.sub.10O.sub.17:Eu, Mn; CaAl.sub.12O.sub.19:Eu; SrAl.sub.12O.sub.19:Eu; SrMgAl.sub.10O.sub.17:Eu; Ba(Al.sub.2O.sub.3).sub.6:Eu; (Ba,Sr)MgAl.sub.10O.sub.17:Eu, Mn; CaAl.sub.2O.sub.4:Eu, Nd; SrAl.sub.2O.sub.4:Eu, Dy; Sr.sub.4Al.sub.14O.sub.25:Eu, Dy as well as mixtures thereof. 7. Silicates: for instance BaSrMgSi.sub.2O.sub.7:Eu; Ba.sub.2MgSiO.sub.7:Eu; BaMg.sub.2Si.sub.2O.sub.7:Eu; CaMgSi.sub.2O.sub.6:Eu; SrBaSiO.sub.4:Eu; Sr.sub.2Si.sub.3O.sub.8.SrCl.sub.2:Eu; Ba.sub.2SiO.sub.4Br.sub.6:Eu; Ba.sub.2SiO.sub.4Cl.sub.6:Eu; Ca.sub.2MgSi.sub.2O.sub.7:Eu; CaAl.sub.2Si.sub.2O.sub.8:Eu; Ca.sub.1.5Sr.sub.0.5MgSi.sub.2O.sub.7:Eu; (Ca,Sr).sub.2MgSi.sub.2O.sub.7:Eu, Sr.sub.2LiSiO.sub.4F:Eu as well as mixtures thereof. 8. Tungstates and molybdates: for instance X.sub.3WO.sub.6 (X=Mg, Ca, Sr, Ba); X.sub.2WO.sub.4 (X=Li, Na, K, Rb, Cs), XMoO.sub.4 (X=Mg, Ca, Sr, Ba) as well as polymolybdates oder polytungstates or the salts of the corresponding hetero- or isopolyacids as well as mixtures thereof. 9. Germanates: e.g. Zn.sub.2GeO.sub.4 10. Moreover the following classes: ALnO.sub.2:Yb, Er (A=Li, Na; Ln=Gd, Y, Lu); LnAO.sub.4:Yb,Er (Ln=La, Y; A=P, V,As, Nb) ; Ca.sub.3Al.sub.2Ge.sub.3O.sub.12:Er; Gd.sub.2O.sub.2S:Yb, Er; La.sub.2SYb, Er as well as mixtures thereof.

(104) Transferring micelle preparations from an aqueous environment into a non-aqueous environment normally results in a deterioration of structural integrity and physico chemical properties of the micelle, because the entropic gain from the release of water molecules during micelle formation is lost. As an example, aqueous micelle preparations of a surfactant, e.g. sodium dodecyl sulfate (SDS) decompose, when transferred to methanol, or even do not constitute when SDS is mixed with methanol (G. D. Parfitt and J. A. Wood Light scattering of sodium dodecyl sulphate in methanol-water mixtures, Kolloid-Zeitschrift and Zeitschrift fr Polymere, January 1969, Volume 229, Issue 1, pp 55-60).

(105) It has been demonstrated that block copolymers comprised of at least two polymer blocks of different compatibility with respect to a given solvent, may form micellar structures in non-aqueous solvents (S. Hou et al., Langmuir 2003, 19, 2485-2490; B.C. Kumi et al., Journal of Colloid and Interface Science 386 (2012) 212-217; P. Alexandridis, Macromolecules 1998, 31, 6935-6942; ibid. Macromolecules 2000, 33, 3382-3391). However, said preparations of polymer micelles in non-aqueous solution showed a lower aggregation number and higher critical micelle concentration (CMC) compared to the aqueous preparation, and hence showed reduced stability. Most obviously, said polymer micelles did not contain any particles.

(106) Surprisingly, application of the invention described herein, revealed that the physico chemical properties of said nanoparticles were much better maintained than in an aqueous preparation, as could be demonstrated by preparation of a stable non-aqueous composition of otherwise highly damageable CdSeTe/CdS nanoparticles. Moreover, said composition in a water miscible solvent could be freely transferred into aqueous solutions without compromising the physico-chemical properties, i.e. fluorescence.

(107) Hence, the present invention describes compositions of micellar enclosed nanoparticles in non-aqueous solvents, methods of preparation and uses, which avoid the shortcomings of aqueous compositions mentioned above.

(108) Furthermore, it was found that micellar encapsulated nanoparticles can be transferred from an aqueous preparation of said nanoparticles into various non-aqueous solvents, completely maintaining structural integrity and physico chemical properties of said nanoparticles, hence fully leveraging on the benefits offered by non-aqueous preparations as pointed out above.

(109) First Aspect of the Present Invention

(110) In a first aspect of the present invention, the micellar encapsulation of hydrophobic nanoparticles may be achieved under completely anhydrous conditions, using combinations of said nanoparticles and said micelle forming unimers in mixtures of intermiscible non-aqueous solvents.

(111) In other words, a method is provided for providing a composition comprising at least one micelle in non-aqueous solution, wherein the micelle encapsulates one or more nanoparticle(s), wherein the micelle encapsulating the one or more nanoparticle(s) is or are formed in the non-aqueous solution.

(112) For instance, nanoparticles, in particular said nanoparticles above, may be co-dissolved with said micelle forming unimers in at least one intermiscible non-aqueous solvent compatible with the hydrophobic domains of said micelle forming unimers and transferred into at least one non-aqueous solvent of reduced compatibility with regard to said unimers.

(113) The micelle forming elements or unimers may include block co-polymers (e.g. amphiphilic block co-polymers), comprised of at least two different polymer blocks of different compatibility towards solvents.

(114) Said selected copolymers may be comprised of the elements selected from C, H, O, N, Si, S, P, X, wherein X represents a halogen selected from the elements F, Cl and Br.

(115) Said block co-polymers may be comprised of about 1 to about 99% by mole fraction of a polymer block incompatible with the other polymer blocks said block co-polymer is comprised of.

(116) An example, which should in no way construed to limit the invention, is a block co-polymer comprised of a block of polystyrene (PS) and polyisoprene (PI), which displays said micelle forming properties, due to the incompatibility of said PS-and PI-block in non-aqueous solvents.

(117) Said block co-polymers may be linear, branched, brush-like, star-shaped or dendritic.

(118) Said block co-polymers may display amphiphilic properties. Furthermore, said amphiphilic block copolymers may be comprised of about 1 to about 99% by mole fraction of hydrophobic segments and about 99 to about 1 mole fraction % of hydrophilic segments.

(119) Hydrophilic segments may be selected from the group consisting of polyethers in particular polyethylene glycol, which is synonymously called polyethylene oxide, polyacetals, polyamides, polylactones, polyimides including poly-2-oxazolines, polyamines, including polyethylene imine and polyallyl amine polylactams, polymaleic acids, polymaleic amides, polymaleic imides, hydrophilic polymaleic esters, polyacrylic acid, polyacrylic amides, hydrophilic polyacrylates, polymethacrylic acid, polymethacrylic amides, hydrophilic polymethacrylates like poly(hydroxyethyl methacrylate), poly-N-vinyl-2-pyrrolidone, poly-N-vinyl-2-piperidone, poly-N-vinyl-2-caprolactam, poly-N-vinyl-3methyl-2-caprolactam, poly-N-vinyl-3methyl-2-piperidone, poly-N-vinyl-4-methyl-2-piperidone, poly-N-vinyl-4-methyl-2-caprolactam, poly-N-vinyl-3-ethyl-2-pyrrolidone, and poly-N-vinyl-4,5-dimethyl-2-pyrrolidone, polyvinylimidazole, poly-N-N-dimethylacrylamide, poly-N-vinyl-N-methylacetamide, polyvinyl alcohol, polyurethanes, polyureas, and biopolymers including amino acid polymers like polylysines, polyhistidines, polyglycines, polyglutamic acids, polyaspartic acids and the corresponding esters and amides, polysaccharides e.g. polydextranes, modified polysaccharides e.g. polycarboxydextranes, carboxymethylcelluloses, hydroxyalkylcelluloses, alginates, pectines and combinations and copolymers thereof. Said polymers may optionally contain charged moieties, e.g. anionic, cationic or zwitterionic groups.

(120) Said hydrophobic segments may be selected from the group consisting of polyolefines, e.g. polyethylene, polypropylene, polybutylene, polyisobutylene, polybutadiene, polyisoprene, polystyrene, cross-linked polystyrenes, poly-p-xylylenes, polyvinylchloride, polyvinylidenechloride, polytetrafluorethylene, polyacrylates, polymethacrylates, e.g. poly(meth)alkylacrlyates, polyethers, e.g. polyoxymethylene, polyphenyleneoxide, polyetheretherketone, polyesters, e.g. polyethyleneterephthalate, polybutyleneterephthalate, polylactic acid, polyglycolic acid, polyvinyl acetate, polyamides, e.g. polyamide-6, polyamide-6.6, polyimide-6.10, polyamide-6.12, polyamide-11, polyamide-12, hydrophobic biopolymers, including natural rubbers, polyurethanes, polyureas, polyphosphazenes and polydialkylsiloxanes, e.g. polydimethylsiloxanes.

(121) Suitable amphiphilic block co-polymers may include, but are not limited to, polystyrene-block-polyethylene oxide, polystyrene-block-poly-L-lactide, polyglycolide-block-polyethylene oxide, polylactide-glycolide-block-polyethylene oxide, polystyrene-block-polymethacrylic acid, polystyrene-block-polyacrylic acid, poly(butadiene)-block-poly(ethylene oxide) poly(isoprene)-block-poly(2-vinpyridine), poly(isoprene)-block-poly(ethylene oxide), poly(methyl methacrylate)-block-poly(2-hydroxyethyl methacrylate), polystyrene-block-poly(2-vinylpyridine), polystyrene-block-poly(4-vinpyridine), poly(methacrylic acid)-block-poly(butyl acrylate), poly(ethylene glycol) methyl ether-block-poly(D,L lactide), poly(ethylene glycol)-block-poly(D,L lactide), poly(ethylene glycol) methyl ether-block-poly(D,L lactide)-block-decane, poly(ethylene glycol) -block-poly(D,L lactide)-block-decane, poly(ethylene glycol)-block-polylactide methyl ether ,poly(L-lactide)-block-poly(ethylene glycol)methyl ether polylactide, poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolide), poly(ethylene glycol)-block-poly(lactide-co-glycolide), poly(ethylene glycol)-block-poly(-caprolactone) methyl ether, poly(ethylene glycol)-block-poly(-caprolactone), polyethylene-block-poly(ethylene glycol), polyethylene-block-poly(ethylene glycol) methyl ether, poly(styrene)-block-poly(ethylene glycol), poly(styrene)-block-poly(ethylene glycol) methyl ether, poly(ethylene glycol) -block-polypropylene glycol (Poloxamer), polyoxyethylen(20)-sorbitan-monooleate (TWEEN 80).

(122) Examples of Y-shaped block copolymers, optionally comprised of two optionally different hydrophilic and one hydrophobic polymer chain or vice versa, include, but are not limited to (PI).sub.2-b-PEO, PI-b-(PEO).sub.2, PS.sub.2-b-PEO, (polystyrene).sub.2-block-poly(ethylene oxide), (polystyrene).sub.2-block-poly(L-lactide), (polystyrene).sub.2-block-poly(methyl methacrylate), (polystyrene).sub.2 -block-poly(tert-butyl methacrylate), polystyrene-block-[poly(tert-butyl acrylate)].sub.2, polystyrene-block-[poly(acrylic acid)].sub.2, polystyrene-block-[poly(ethylene oxide)].sub.2.

(123) Examples of branched block copolymers may include, but are not limited to poly(ethylene oxide)-block-polylactide, 4-arm poly(ethylene oxide), poly(ethylene oxide)-block-polycaprolactone, 4-arm poly(ethylene oxide).

(124) Examples of dendritic amphiphilic block copolymers are given in the literature, e.g. Virgil Percec, et al. Self-Assembly of Janus Dendrimers into Uniform Dendrimersomes and Other Complex Architectures, Science 328, 1009 (2010).

(125) The micellar encapsulated nanoparticles thus obtained can be used directly or optionally further processed by crosslinking said micelle forming polymers using processes described in the literature, in particular patent application WO 2010/100108 A1. Crosslinking includes, but is not limited to radical induced crosslinking, radiation induced crosslinking, e.g. by light of different wavelengths like x-ray, UV, visible or infrared, thermal crosslinking or chemical crosslinking, e.g. ring opening of multiple epoxide moieties with nucleophiles, e.g. amines, in particular polyamines providing more than one amino group, or vulcanization processes with sulfur containing compounds, e.g. disulfur dichloride and the like.

(126) As a result, colloidal compositions of perfectly dispersed nanoparticles in micelles in a non-aqueous environment are obtained. The generation of micelles, e.g. in methanol was evident from the fact that said nanoparticles without micelle forming additives were completely insoluble in methanol. Micelle formation was further proven via DLS measurements (see FIG. 1). FIG. 1 shows the DLS-Data of CdSeTe/CdS (core/shell) nanoparticles after the encapsulation in polymeric micelles in methanol and after the transfer into water. Said nanoparticles still display virtually identical properties like the native nanoparticles, e.g. fluorescent semiconductor nanoparticles still display virtually identical optical properties like the native nanoparticles (see e.g. FIG. 2 (absorption spectrum (left) and emission spectrum (right)).

(127) Suitable non-aqueous solvents may include, but are not restricted to unbranched, branched or cyclic alkanes, optionally bearing one or more halogen atoms, like pentane, hexane, iso-hexanes, cyclopentane, cyclohexane, dichloromethane, chloroform, 1,1-dichloroethane, or 1,2-dichloroethane; halogenated alkenes, like 1,1-dichloroethene or all isomers of 1,2-dichloroethene; optionally alkylated aromatic or heteroaromatic solvents, optionally bearing one or more halogen atoms, e.g., benzene, toluene, ethyl benzene, xylenes, chloro benzene, pyridine, collidines or lutidines; unbranched, branched or cyclic ethers, like diethyl ether, diisopropyl ether, methyl t-butyl ether, tetrahydrofuran or 1,4-dioxane; carbonyl compounds, in particular ketones like acetone, methyl ethyl ketone, cylopentanone or cyclohexanone; carboxylic acid esters, e.g. ethyl acetate and the like, unbranched or branched C.sub.1-C.sub.5 alkanols, like methanol, ethanol, n-propanol, i-propanol, or all isomers of butanol and pentanol; polyols, like ethylene glycol, 1,2-propanediol, 1,3-propanediol, glycerol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 1,2,4-butanetriol, diethylene glycol (DEG); ethers of polyols, e.g. 2-methoxyethanol, bis-(2-methoxyethyl) ether (diglyme); esters of polyols, e.g. diacetoxyethane; carboxylic acids, like formic acid, acetic acid, propanoic acid, lactic acid; dialkyl sulfoxides, e.g. dimethyl sulfoxide (DMSO); carboxylic acid amides, e.g. formamide, acetamide, N-methyl formamide, N-methyl acetamide, N,N-dimethyl formamide (DMF), N,N-dimethyl acetamide (DMA), or N-methyl pyrrolidone (NMP); phosphoric acid amides e.g. hexamethyl phosphoric acid triamide (HMPA); urea based solvents like 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU), 1,3-dimethyl-2-imidazolidinone (DMI), or tetramethyl urea (TEMUR); lower alkylnitriles like acetonitrile, propionitrile or butyronitrile; ionic liquids, e.g. ammonium-based ionic liquids; imidazolium based ionic liquids like 1-allyl-3methylimidazolium, 1-benzyl-3methylimidazolium, 1-butyl-2,3-dimethylimidazolium, 1-butyl-3methylimidazolium, 1-decyl-3-methylimidazolium, 1,3-didecyl-2-methylimidazolium, 1,3-dimethylimidazolium, 1,2-dimethyl-3-propylimidazolium, 1-dodecyl-3-methylimidazolium, 1-ethyl-2,3-dimethylimidazolium, 1-ethyl-3methylimidazolium, 1-hexadecyl-3-methylimidazolium, 1-hexyl-3methylimidazolium, 1-(2-hydroxyethyl)-3-methylimidazolium, 1-methyl-3octadecylimidazolium, 1-methyl-3octylimidazolium, 1-methyl-3-propylimidazolium salts, preferably with complex anions like e.g. tetrafluoroborate; piperidinium-based ionic liquids, like 1-butyl-1-methylpiperidinium, 1-methyl-1-propylpiperidinium; pyridinium-based ionic liquids, 1-alkyl-2-methylpyridinium, 1-alkyl-3-methylpyridinium, 1-alkyl-4-methylpyridinium, 1-butylpyridinium, 1-propylpyridinium, 1-ethylpyridinium, 1-hexylpyridinium salts, preferably with complex anions like e.g. tetrafluoroborate; pyrrolidinium-based ionic liquids, e.g. 1-butyl-1-methylpyrrolidinium, 1-ethyl-1-methylpyrrolidinium, 1-methyl-1-propylpyrrolidinium salts, preferably with complex anions, and combinations or combinations of mixtures thereof.

(128) In particular, the non-aqueous solution may comprise at least partially a water miscible non-aqueous solvent and/or wherein the non-aqueous solution comprises a non-aqueous solvent, which does not form azeotropic phases with water.

(129) Furthermore, the water miscible non-aqueous solvent may have a higher boiling point than water.

(130) Second Aspect of the Present Invention

(131) In a second aspect of the invention, nanoparticles may be enclosed in micelles in an aqueous preparation and the micelle surface is subsequently crosslinked by known chemical methods, e.g. condensation, radical induced polymerization and the like, as comprehensively described in patent applications WO 2012/001012 A2 and WO 2010/100108 A1. Said aqueous preparation of nanoparticles enclosed in preferably crosslinked micelles can be successively transferred in a non-aqueous solution, provided that the non-aqueous solvent is at least partially miscible with the aqueous solution, herefrom the water is removed.

(132) In other words, a method is provided for providing a composition comprising at least one micelle in non-aqueous solution, wherein the micelle encapsulates one or more nanoparticle(s), wherein in a first step the micelle encapsulating the one or more nanoparticle(s) is formed in a first solution being different from the non-aqueous solution and wherein in a second step the first solution is replaced by the non-aqueous solution.

(133) Then the aqueous preparation of said nanoparticles in cross-linked micelles may be mixed with at least one water miscible non-aqueous solvent with a higher boiling point than the aqueous preparation of said nanoparticles.

(134) Water miscible within the scope of the invention means that a solvent forms a homogenous phase with water, containing at least 1% per volume of water.

(135) Preferably said non-aqueous solvents may not form azeotropic phases with water.

(136) The water may be subsequently removed, preferably by distillation at or below atmospheric pressure.

(137) Suitable non-aqueous solvents may include, but are not restricted to dialkyl sulfoxides, e.g. dimethyl sulfoxide (DMSO); carboxylic acid amides, e.g. formamide, acetamide, N-methyl formamide, N-methyl acetamide, N,N-dimethyl formamide (DMF), N,N-dimethyl acetamide (DMA), or N-methyl pyrrolidone (NMP); phosphoric acid amides e.g. hexamethyl phosphoric acid triamide (HMPA); urea based solvents like 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU), 1,3-dimethyl-2-imidazolidinone (DMI), or tetramethyl urea (TEMUR); 1,4-dioxane; higher alkanols, e.g. 1-butanol; polyols, like ethylene glycol, 1,2-propanediol, 1,3-propanediol, glycerol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 1,2,4-butanetriol, diethylene glycol (DEG); ethers of polyols, e.g. 2-methoxyethanol, bis-(2-methoxyethyl) ether (diglyme); esters of polyols, e.g. diacetoxyethane; carboxylic acids, like formic acid, acetic acid, propanoic acid, lactic acid; primary, secondary and tertiary amines, diamines or triamines bearing branched, unbranched or cyclic alky groups on the nitrogen atoms and being liquids at ambient temperature; cyclic or bicylic amines, optionally bearing further heteroatoms, selected from N, O, or S in the ring, like piperidine, N-methyl piperidine, morpholine, N-methyl morpholine; annelated amidine bases, like 1,5-diazabicyclo(4.3.0)non-5-ene (DBN) or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU); optionally alkylated pyridines, e.g. pyridine, all isomers of picoline, lutidine and collidine; C1-2 nitro alkanes; ionic liquids, e.g. ammonium-based ionic liquids; imidazolium based ionic liquids like 1-allyl-3methylimidazolium, 1-benzyl-3methylimidazolium, 1-butyl-2,3-dimethylimidazolium, 1-butyl-3methylimidazolium, 1-decyl-3-methylimidazolium, 1,3-didecyl-2-methylimidazolium, 1,3-dimethylimidazolium, 1,2-dimethyl-3-propylimidazolium, 1-dodecyl-3-methylimidazolium, 1-ethyl-2,3-dimethylimidazolium, 1-ethyl-3methylimidazolium, 1-hexadecyl-3-methylimidazolium, 1-hexyl-3methylimidazolium, 1-(2-hydroxyethyl)-3-methylimidazolium, 1-methyl-3octadecylimidazolium, 1-methyl-3octylimidazolium, 1-methyl-3-propylimidazolium salts, preferably with complex anions like e.g. tetrafluoroborate; piperidinium-based ionic liquids, like 1-butyl-1-methylpiperidinium, 1-methyl-1-propylpiperidinium; pyridinium-based ionic liquids, 1-alkyl-2-methylpyridinium, 1-alkyl-3-methylpyridinium, 1-alkyl-4-methylpyridinium, 1-butylpyridinium, 1-propylpyridinium, 1-ethylpyridinium, 1-hexylpyridinium salts, preferably with complex anions like e.g. tetrafluoroborate; pyrrolidinium-based ionic liquids, e.g. 1-butyl-1-methylpyrrolidinium, 1-ethyl-1-methylpyrrolidinium, 1-methyl-1-propylpyrrolidinium salts, preferably with complex anions, which in combination with the cation renders the ionic liquid water miscible, like e.g. tetrafluoroborate, or mixtures of said solvents.

(138) Some of the mentioned solvents may exist as isomers, geometrical isomers, stereoisomers, e.g. diastereomers or enantiomers, meso compounds or racemates. It should be understood that all isomers of said non-aqueous solvents are in the scope of the invention.

(139) Some of the above mentioned solvents may be toxic to cell organelles, living cells, living tissues, organs or organisms. In some cases, e.g. antimicrobial activity may be a desirable feature of a composition of micelle encapsulated nanoparticles.

(140) However, in general, said compositions of micelle encapsulated nanoparticles in solvents with minimal or no toxic side effects for the desired application are preferred.

(141) Third Aspect of the Present Invention

(142) In a third aspect of the invention, residual water may be removed from the micelle encapsulated nanoparticle preparation in non-aqueous solutions solely or supportingly by chemical or physical processes (see e.g. the second aspect of the present invention above), well known in the art. These processes include, but are not limited to, azeotropic removal of water, e.g. using solvents like toluene; diffusion or permeation across porous or non-porous membranes, including dialysis and pervaporation; or chemical drying using water-absorbing materials, including but not limited to, inorganic salts, like Na.sub.2SO.sub.4, MgSO.sub.4, or CaCl.sub.2, or minerals like e.g. bentonite; or polymers, in particular superabsorbent polymers (SAPs), e.g. polyacrylate- or cellulose based polymers and the like.

(143) Fourth Aspect of the Present Invention

(144) In a fourth aspect of the invention, a non-aqueous preparation of nanoparticles enclosed in preferably crosslinked micelles may be prepared from said aqueous preparation of nanoparticles enclosed in preferably crosslinked micelles by evaporating said aqueous preparation to dryness and dissolution of the residue in a suitable solvent, including the solvents mentioned above.

(145) In general, all above disclosed compositions may be used for a biological event or biological application.

(146) The term biological event or biological application has to be understood in the broadest sense.

(147) In particular, the term biological event inter alia also covers the wanted or unwanted transfection of cells or the wanted or unwanted introduction of micelles of the composition into cells. Also, the conjugation to cells, the labeling of cells, cell compartments or subcellular organelles, the targeted or non-targeted transfer of biomolecules into the cell, cell compartments or subcellular organelles, the triggered or non-triggered release of the transferred biomolecules and the detection of a response shall be covered.

(148) The term biological application also covers the use of the composition or parts of the composition for marking of biological liquids, organic components and structures, cells or the like. Said biological applications include, but are not limited to, the analytic or diagnostic detection of biomolecules, biological targets or event factors, the detection of target molecules form analytes by e.g. nucleic acid NP compositions like e.g. single stranded DNA-NP conjugates, single stranded RNA-NP conjugates, single stranded siRNA-NP conjugates, single stranded LNA-NP conjugates, double-stranded or multi-stranded variations thereof and combined products to be used in nucleic acid assays, like e.g. polymerase chain reaction (PCR), quantitative PCR, reverse transcriptase PCR, quantitative reverse transcriptase PCR, digital PCR or Helicase-dependent DNA amplification (HAD), isothermal HAD and the like, the detection of target molecules form analytes by e.g. aminoacid NP compositions like e.g. protein-NP conjugates, peptide-NP comjugates, antibody-NP comjugates, antibodylike-NP conjugates, aptamer-NP conjugates, by molecular detection methods like e.g. Western Blot, ELISA, DOT-Blot, Slot-Blot, Northern Blot, Southern Blot techniques and variations thereof. An example of blot variation is the NP composition optimized fast version, which only requires the incubation of immobilized target DNA with NP DNA molecular probes generated e.g. as described in example 10 (DNA Conjugation FIG. 4), and the analysis or detection of analytes in competition assays (FIG. 5).

EXAMPLE 1

(149) General Procedure for the Preparation of Micelle Encapsulated Nanoparticles in Methanol

(150) In the typical procedure, 5 nmol of nanoparticles in chloroform are mixed with 2.5 mol of the diblock copolymer poly(isoprene-block-ethylene oxide) (PI-b-PEO; M.sub.N=13700 g/mol) and diluted to a total volume of 500 L with chloroform. Subsequently the homogeneous nanoparticle/polymer solution is quickly injected into a stirred solution of 5000 L methanol. The resulting solution is stirred for a minute. The encapsulation of the nanoparticles within the micellular core region can be concluded from the fact, that all used nanoparticles bear hydrophobic surface ligands, which makes the native particles insoluble in methanol. However, after the micelle encapsulation process a homogenous solution of the nanoparticles is obtained, showing comparative characteristics as the native solutions.

EXAMPLE 2

(151) Non-aqueous Composition of Micelle Encapsulated Fluorescent Trioctylamine-coated CdSeTe/CdS Nanoparticles in Methanol

(152) 2 nmol of trioctylamine-coated CdSeTe/CdS core/shell nanoparticles and 2.4 mol (1200 fold excess) of polyisoprene-b-polyethyleneoxide diblock copolymer (PI-b-PEO) with an average mol weight of 13700 g/mol and a PI/PEO weight ratio of 1:2 were dissolved in 200 L of chloroform and then quickly injected into 2 mL of methanol. A clear, slightly colored solution was obtained, indicating that the starting material, which is completely insoluble in methanol, was successfully encapsulated in PI-b-PEO micelles. Accordingly, dynamic light scattering (DLS) indicated micelles with diameters of about 20 to 30 nm. (FIG. 1 and FIG. 2)

EXAMPLE 3

(153) Non-aqueous Composition of Micelle Encapsulated Fluorescent Octadecylamine-coated CdSeTe/CdS Nanoparticles

(154) 2 nmol of octadecylamine-coated CdSeTe/CdS core/shell nanoparticles were incubated with 1.2 mol (600 fold excess) of diethylene triamine functionalized polyisoprene (PI-DETA; 3000 g/mol) in chloroform and then precipitated by the addition of ethanol. Subsequently, the PI-DETA-coated nanoparticles and 2.4 mol (1200 fold excess) of the PI-b-PEO diblock copolymer (13700 g/mol) were dissolved in 200 L of chloroform and then quickly injected into 2 mL of methanol. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS), indicating micelles with diameters of about 20 to 30 nm.

EXAMPLE 4

(155) Non-aqueous Composition of Micelle Encapsulated Fluorescent TOP/TOPO-coated CdSe/CdS/ZnS Particles

(156) Similar to Example 3, 5 nmol of TOP/TO(PO-coated CdSe/CdS/ZnS core/shell/shell nanoparticles were precoated with 3.0 mol (600 fold excess) of PI-DETA (3000 g/mol) and then precipitated by the addition of ethanol. Subsequently, the particles and 1.5 mol (300 fold excess) of the PI-b-PEO diblock copolymer (13700 g/mol) were dissolved in 200 L chloroform and then quickly injected into 2 mL methanol. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS), indicating micelles with diameters of about 20 to 30 nm. (Spectral properties FIG. 3)

EXAMPLE 5

(157) Transfer of Micellar Encapsulated Nanoparticles from Methanol to Water

(158) In a subsequent step the micellar encapsulated nanoparticles are transferred from methanol to water.

(159) FIG. 1 shows DLS-Data for micellar encapsulated nanoparticles in methanol (dashed line) and water (solid line).

(160) First, most of the solvents were removed by rotary evaporation. The concentrated solution was injected into 10 mL of water, resulting in a clear, slightly colored solution. Minor methanol impurities were removed by several washing steps with water, using centrifugal concentrators (Sartorius VivaSpin 20). The DLS data show slightly increased hydrodynamic diameters for micelles in water compared to micelles in methanol, due to swelling of the PEO block in water.

EXAMPLE 6

(161) a) Composition of Micelle Encapsulated Fluorescent Quantum Dots in Aqueous Solution

(162) An aqueous composition of fluorescent quantum dot-rods was prepared according to: Jan Niehaus, Sren Becker, Christian Schmidtke, Daniel Ness, Katja Werner, Horst Weller, Water-soluble nanoparticles as an universal imaging tool, From Nanotech Conference & Expo 2012: An Interdisciplinary Integrative Forum on Nanotechnology, Microtechnology, Biotechnology and Cleantechnology, Santa Clara, Calif., United States, Jun. 18-21, 2012, 1, 413-416 (2012), starting from cadmium selenide/cadmium sulfide CANdots quantum rods purchased from STREM Inc (CdSe/CdS elongated core/shell; CAS Number: 1306-24-7; MDL Number: MFCD00171259; 590 nm peak emission). Also, the methods as disclosed by WO 2012/001012 A2 and WO 2010/100108 A1 can be used and have been used for enabling this possible embodiment.

(163) b) Non-aqueous Composition of Micelle Encapsulated Fluorescent Quantum Dot-rods in rac-1,2-propanediol

(164) 1 mL of a 5 M aqueous micellar preparation of fluorescent quantum dot-rods, according to Example 2 were added to 1 mL of rac-1,2-propanediol (Sigma-Aldrich, 39,803-9, Lot. 04430JJ-249) in a round bottom flask. Water was removed by rotary evaporation (70 hPa, 60 C. bath temperature) to a residual volume of 950 L in the flask.

(165) A sample of 50 L of the preparation of fluorescent quantum dot-rods in rac-1,2-propanediol were diluted with distilled water to an overall volume of 3 mL. A clear solution was obtained without any precipitation, even after extended storage. From this solution an UV spectrum was recorded and the hydrodynamic diameter was estimated by dynamic light scattering (DLS). No differences compared to a corresponding dilution of the starting composition of Example 2 in water were detectable.

EXAMPLE 7

(166) Non-aqueous Composition of Micelle Encapsulated Fluorescent Quantum Dot-rods in rac-1,3-butanediol

(167) The experiment of Example 6 was repeated, replacing rac-1,2-propanediol with rac-1,3-butanediol. A dilution of this composition in water showed no differences compared to a dilution of the starting aqueous composition of Example 2.

EXAMPLE 8

(168) Conjugation of DNA to Micelle Encapsulated Fluorescent Quantum Dot-rods in rac-1,2-propanediol

(169) An aliquot of 10 L of the composition obtained in Example 6 was mixed with 10.8 L of a 92 M stock solution of 5 -thiol terminated DNA (33 mer) and adjusted to 50 L by addition of rac-1,2-propanediol. The sample was heated to 50 C. for 2 h in a heating block. An aliquot of 5 L was kept for analysis and the remaining volume was heated to 95 C. for 15 min. The samples were analyzed via gel electrophoresis on agarose. Aliquots of 2.5 L of each sample were diluted with 22.5 L distilled water and 5 L gel loading buffer was added. Aliquots of 25 L of this mixture were applied to a 0.8% agarose gel. After processing the electrophoresis by applying 90 V voltage for 1 h, samples were visualized by UV excitation of the quantum dot-rods. Successful conjugation was indicated by progression towards the anode due to the negative charge of the conjugated nanoparticles, whereas the unconjugated controls in 1,2-propanediol moved to the cathode. It was revealed that the conjugation of DNA is much more efficient in non-aqueous solution than in aqueous solution, most likely due to preservation of the stretched strand, whereas in aqueous solution a secondary structure will be taken to preserve hydrophobic bases from the environment. Heating to 95 C. was even more efficient than heating to 50 C. (FIG. 4).

(170) For purification the nanoparticle-DNA conjugates were transferred into an ultrafiltration unit and diluted with 5 mL of 0.5 PBS. After centrifugation at 1200*g for 20 min the supernatant was discarded and centrifugation continued until a residual volume of 350 L was achieved.

EXAMPLE 9

(171) Conjugation of DNA to Micelle Encapsulated Fluorescent Quantum Dot-rods in rac-1,3-butanediol

(172) Performing the experiment of Example 8 with a composition of encapsulated fluorescent quantum dot-rods in rac-1,3-butanediol provided identical results (FIG. 4)

EXAMPLE 10

(173) Functional Proof of Fluorescent Quantum Dot-rod-DNA Conjugates

(174) Aliquots of 1 L of a 45 M NeutrAvidin stock solution (3.0 mg/ml) were spotted on stripes of nitrocellulose membrane and dried. Membranes were incubated with a solution of biotinylated complementary and biotinylated non-complementary DNA (cDNA). The membranes were incubated with fluorescent quantum dot-rod-DNA conjugates as described in example 8 and 9. An accumulation of the fluorescent quantum dot-rods-DNA conjuugates was clearly detectable only on spots providing the complementary DNA but not on the control or on the blank membrane (FIG. 5).

EXAMPLE 11

(175) a) Composition of Micelle Encapsulated Fluorescent Quantum Dots in Aqueous Solution

(176) An aqueous composition of fluorescent quantum dots was prepared similarly to example 6a starting from cadmium selenide/zinc selenide/zinc sulfide CANdots Series A org quantum dots commercially available from CAN GmbH, Germany (CdSe/ZnSe/ZnS core/shell/shell; 610 nm peak emission).

(177) b) Conjugation of Biotin to Micelle Encapsulated Fluorescent Quantum Dots in 10% DMSO

(178) 29 mL of a 0.5 M aqueous micellar preparation of fluorescent quantum dots, according to Example 6a were added to 3 mL DMSO Sigma-Aldrich, (CAS 27,685-5) supplemented with 50 M NHS-LC-LC-Biotin (N-Succinimidyl N-[6-(biotinylamino)caproyl]-6-aminocaproate, 6-(Biotinamidocaproylamido)caproic acid N-hydroxysuccinimide ester, AppliChem, Product: A7856,0050, MW 567.70 g/mol, CAS: 89889-52-1) in a 50 mL screw capped glass vial. The biotinylation reaction processed via NHS reaction on NH2-exposing micelle components upon incubation for 1.5 h at 40 C.

(179) For purification the biotinylated nanoparticle micelles were transferred into two ultrafiltration units and concentrated by centrifugation in a Sorvall RC6+high-speed centrifuge equipped with rotor 42, at 4.000 rpm at approx. 40 C. for 30 min. From each concentrator 5 mL sample was recovered and the pooled concentrate was diluted with 10 mL 1 PBS (1:10 dilution in ultrapure water from a 10 stock prepared from 100 g NaCl, 2.5 g KCl, 18 g Na.sub.2HPO.sub.4.2H.sub.2O, 3 g KH.sub.2PO.sub.4 in 1 L of distilled water) supplemented with 0.0025% lithium-azide (CAS Number 19597-69-4). The final 10 mL sample was directly used for access NeutrAvidin binding a subsequent binding of biotinylated biomolecules to remaining free biotin binding sites on the thus NeutrAvidin functionalized micelle encapsulated fluorescent quantum dots.

EXAMPLE 12

(180) Composition of Micelle Encapsulated, Fluorescent, Trioctylamine Coated, 6 nm CdSeTe/CdS Nanoparticles in Methanol

(181) 2 nmol of trioctylamine-coated CdSeTe/CdS core/shell nanoparticles and 2.4 mol (1200 fold excess) of poly(isoprene-b-ethylene oxide) diblock copolymer (PI-b-PEO) with an average mol weight of 13700 g/mol and a PI/PEO block weight ratio of 1:2 were dissolved in 200 L of chloroform and then quickly injected into 2 mL of methanol. A clear, slightly colored solution was obtained, indicating that the starting material, which is completely insoluble in methanol, was successfully encapsulated in PI-b-PEO micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 6). As can be seen from the spectroscopic measurements (FIG. 7 and FIG. 8), the absorption and emission properties of the nanoparticles are virtually maintained.

EXAMPLE 13

(182) Composition of Micelle Encapsulated, Fluorescent, Octadecylamine Coated, 6 nm CdSeTe/CdS Nanoparticles in Methanol

(183) 2 nmol of octadecylamine-coated CdSeTe/CdS core/shell nanoparticles were incubated with 1.2 mol (600 fold excess) of diethylene triamine functionalized polyisoprene (PI-DETA; 3000 g/mol) in chloroform and then precipitated by the addition of ethanol. Subsequently, the PI-DETA-coated nanoparticles and 2.4 mol (1200 fold excess) of the PI-b-PEO diblock copolymer (13700 g/mol) were dissolved in 200 L of chloroform and then quickly injected into 2 mL of methanol. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 9). The absorption and emission properties of the nanoparticles are displayed in FIG. 10 and FIG. 11.

EXAMPLE 14

(184) Composition of Micelle Encapsulated, Fluorescent, Trioctylphosphine/Trioctylphosphine Oxide Coated, 6 nm CdSe/CdS/ZnS Nanoparticles in Methanol

(185) 2.5 nmol of trioctylphosphine/trioctylphosphine oxide (TOP/TOPO) coated CdSe/CdS/ZnS core/shell/shell nanoparticles were precoated with 3.0 mol (600 fold excess) of PI-DETA (3000 g/mol) and then precipitated by the addition of ethanol. Subsequently, the particles and 1.5 mol (300 fold excess) of the PI-b-PEO diblock copolymer (13700 g/mol) were dissolved in 200 L chloroform and then quickly injected into 2 mL methanol. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 12). As can be seen from the spectroscopic measurements (FIG. 13 and FIG. 14), the absorption and emission properties of the nanoparticles are virtually maintained.

EXAMPLE 15

(186) Composition of Micelle Encapsulated, Fluorescent, Oleic Acid/Oleylamine Coated, 8 nm CdSe/CdS Nanoparticles in Methanol

(187) 5 nmol of oleic acid/oleyalamine coated CdSe/CdS core/shell nanoparticles in chloroform were mixed with 2.7 mol (533 fold excess) of PI-b-PEO diblock copolymer (13700 g/mol) and adjusted to a total volume of 500 L with chloroform. Subsequently the solution was quickly injected into 5000 L of methanol and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 15). Absorption and emission properties of the nanoparticles are displayed in FIG. 16 and FIG. 17.

EXAMPLE 16

(188) Composition of Micelle Encapsulated, Fluorescent, Octadecylphosphonic Acide Coated CdSe/CdS Rod-shaped Nanoparticles in Methanol

(189) 5 nmol of octadecylphosphonic acid coated CdSe/CdS rod-shaped nanoparticles (4.8 nm in width; 22.2 nm in length) in chloroform were mixed with 8.8 mol (1750 fold excess) of PI-b-PEO diblock copolymer (13700 g/mol) and adjusted to a total volume of 700 L with chloroform. Subsequently the solution was quickly injected into 5000 L of methanol and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 18). Again, the absorption and emission properties of the nanoparticles are only slightly influenced by the encapsulation process (FIG. 19 and FIG. 20).

EXAMPLE 17

(190) Composition of Micelle Encapsulated, Fluorescent, Octadecylphosphonic Acide Coated CdSe/CdS Rod-shaped Nanoparticles in Acetonitrile

(191) 5 nmol of octadecylphosphonic acid coated CdSe/CdS rod-shaped nanoparticles (4.8 nm in width; 22.2 nm in length) in chloroform were mixed with 8.8 mol (1750 fold excess) of PI-b-PEO diblock copolymer (13700 g/mol) and adjusted to a total volume of 700 L with chloroform. Subsequently the solution was quickly injected into 5000 L of acetonitrile and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 21). Again, the absorption and emission properties of the nanoparticles are virtually maintained (FIG. 22 and FIG. 23).

EXAMPLE 18

(192) Composition of Micelle Encapsulated, Fluorescent, Octadecylphosphonic Acide Coated CdSe/CdS Rod-shaped Nanoparticles in Dimethyl Sulfoxide

(193) 5 nmol of octadecylphosphonic acid coated CdSe/CdS rod-shaped nanoparticles (4.8 nm in width; 22.2 nm in length) in chloroform were mixed with 8.8 mol (1750 fold excess) of PI-b-PEO diblock copolymer (13700 g/mol) and adjusted to a total volume of 700 L with chloroform. Subsequently the solution was quickly injected into 5000 L of Dimethyl sulfoxide and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 24). Again, the absorption and emission properties of the nanoparticles are virtually maintained (FIG. 25 and FIG. 26).

EXAMPLE 19

(194) Composition of Micelle Encapsulated, Fluorescent, Octadecylphosphonic Acide Coated CdSe/CdS Rod-shaped Nanoparticles in N,N-Dimethylformamide

(195) 5 nmol of octadecylphosphonic acid coated CdSe/CdS rod-shaped nanoparticles (4.8 nm in width; 22.2 nm in length) in chloroform were mixed with 8.8 mol (1750 fold excess) of PI-b-PEO diblock copolymer (13700 g/mol) and adjusted to a total volume of 700 L with chloroform. Subsequently the solution was quickly injected into 5000 L of N,N-Dimethylformamide and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 27). The absorption and emission properties are displayed in FIG. 28 and FIG. 29.

EXAMPLE 20

(196) Composition of Micelle Encapsulated, Fluorescent, Octadecylphosphonic Acide Coated CdSe/CdS Rod-shaped Nanoparticles in [BMIM][BF.sub.4]

(197) 1 nmol of octadecylphosphonic acid coated CdSe/CdS rod-shaped nanoparticles (4.8 nm in width; 22.2 nm in length) in chloroform were mixed with 1.8 mol (1750 fold excess) of PI-b-PEO diblock copolymer (13700 g/mol) and adjusted to a total volume of 140 L with chloroform. Subsequently the solution was quickly injected into 1500 L of ionic liquid [BMIM] [BF.sub.4] and stirred for a couple of minutes. The mixing process is very slow due to the high viscosity of the ionic liquid. To fasten this process, the micture was gently heated (ca. 50 C.) while stirring. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 30). The absorption and emission properties of the nanoparticles are displayed in FIG. 31 and FIG. 32.

EXAMPLE 21

(198) Composition of Micelle Encapsulated, Fluorescent, Trioctylphosphine Coated, 5 nm InP/ZnS Nanoparticles in Methanol

(199) 5 nmol of hexdecylamine coated InP/ZnS nanoparticles in toluene were completely dried under a gentle nitrogen flow. The solid residue was dissolved in 300 L chloroform and mixed with 2.5 mol (500 fold excess) of PI-b-PEO. The resulting solution was quickly injected into 2500 L of methanol and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 33). The absorption and emission properties of the nanoparticles are displayed in FIG. 34 and FIG. 35).

EXAMPLE 22

(200) Composition of Micelle Encapsulated, Fluorescent, Hexadecanethiol Coated, 5 nm InP/ZnS Nanoparticles in Methanol

(201) 30 nmol of hexadecylamine coated InP/ZnS nanoparticles were incubated with 90 mol (3000 fold excess) hexadecanethiol. After 24 h of incubation the nanoparticles were precipitated from the solution by the addition of the same volume of methanol. The supernatant was discarded and the pellet was dissolved in 300 L chloroform.

(202) 100 L (approx. 10 nmol) of the hexadecanethiol coated InP/ZnS nanoparticles were mixed with 5 mol (500 fold excess) of PI-b-PEO diblock copolymer (13700 g/mol) and adjusted to a total volume of 500 L with chloroform. Subsequently the solution was quickly injected into 5000 L of methanol and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Again, the absorption and emission properties of the nanoparticles are even improved by the encapsulation process (FIG. 36 and FIG. 37).

EXAMPLE 23

(203) Composition of Micelle Encapsulated, Fluorescent, Trioctylphosphine/Trioctylphosphone Oxide Coated CulnSe/ZnS Nanoparticles in Methanol

(204) 50 L of a stock solution of CuInSe/ZnS nanoparticles in chloroform were mixed with 0.060 g PI-b-PEO (13700 g/mol) and adjusted to a total volume of 250 L with chloroform. Subsequently the solution was quickly injected into 2500 L of methanol and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 38). Again, the absorption and emission properties of the nanoparticles are almost maintained (FIG. 39 and FIG. 40).

EXAMPLE 24

(205) Composition of Micelle Encapsulated, Plasmonic, Dodecanethiol Coated, 4 nm Au Nanoparticles in Methanol

(206) 3 nmol of dodecanethiol coated Au nanoparticles in toluene were dried in a gentle nitrogen flow. The resulting solid was dissolved in 300 L of a solution containing 1.5 mol (500 fold excess) PI-b-PEO in dichloromethane. The resulting solution was quickly injected into 2500 L of methanol and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 41). The plasmon band of the gold nanoparticles is slightly shifted, possibly due to the change of the dielectric environment (FIG. 42).

EXAMPLE 25

(207) Composition of Micelle Encapsulated, Plasmonic, Dodecanethiol Coated, 4 nm Au Nanoparticles in Acetonitrile

(208) 3 nmol of freshly precipitated dodecanethiol coated Au nanoparticles in 300 L CHCl.sub.3 were mixed with 1.5 mol (500 fold excess) PI-b-PEO. The resulting solution was quickly injected into 3000 L of acetonitrile and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 43). The plasmon band of the gold nanoparticles is slightly shifted, possibly due to the change of the dielectric environment (FIG. 44).

EXAMPLE 26

(209) Composition of Micelle Encapsulated, Plasmonic, Dodecanethiol Coated, 4 nm Au Nanoparticles in N,N-dimethylformamide

(210) 3 nmol of freshly precipitated dodecanethiol coated Au nanoparticles in 300 L CHCl.sub.3 were mixed with 1.5 mol (500 fold excess) PI-b-PEO. The resulting solution was quickly injected into 3000 L of N,N-Dimethylformamide (N,N-DMF) and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 45). The plasmon band of the gold nanoparticles is slightly shifted, possibly due to the change of the dielectric environment (FIG. 46).

EXAMPLE 27

(211) Composition of Micelle Encapsulated, Plasmonic, Dodecanethiol Coated, 4 nm Au Nanoparticles in Dimethylsulfoxide

(212) 3 nmol of freshly precipitated dodecanethiol coated Au nanoparticles in 300 L CHCl.sub.3 were mixed with 1.5 mol (500 fold excess) PI-b-PEO. The resulting solution was quickly injected into 3000 L of dimethylsulfoxide (DMSO) and stirred for a minute. The clear, slightly colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 47). The plasmon band of the gold nanoparticles is slightly shifted, possibly due to the change of the dielectric environment (FIG. 48).

EXAMPLE 28

(213) Composition of Micelle Encapsulated, Plasmonic, Dodecanethiol Coated, 4 nm Au Nanoparticles in 1-butyl-3methylimidazolium tetrafluoroborate ([BMIM][BF.sub.4])

(214) 1 nmol of freshly precipitated dodecanethiol coated Au nanoparticles in 200 L CHCl.sub.3 was mixed with 1.5 mol (1500 fold excess) PI-b-PEO. The resulting solution was quickly injected into 1000 L of [BMIM][BR.sub.4] and stirred for a minute. Subsequently the mixture was vigorously stirred and heated carefully to evaporate the remaining CHCl.sub.3. After cooling to room temperature a clear, slightly colored solution was obtained indicating the colloidal dissolution of, the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 49). The plasmon band of the gold nanoparticles is shifted, possibly due to the change of the dielectric environment (FIG. 50).

EXAMPLE 29

(215) Composition of Micelle Encapsulated Magnetic, Oleic Acid, 10 nm Fe.sub.xO.sub.y Nanoparticles in Methanol

(216) 5 nmol Fe.sub.xO.sub.y nanoparticles in toluene were precipitated by the addition of an equal volume of methanol. The cloudy mixture was centrifuged (6000g; 10 min) and the supernatant was discarded. The remaining pellet was dissolved in 500 L chloroform and mixed with 6.9 mol (1390 fold excess) PI-b-PEO. The resulting solution was quickly injected into 5000 L methanol and stirred for a minute. The clear, brownish colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 51).

EXAMPLE 30

(217) Composition of Micelle Encapsulated Magnetic, Oleic Acid, 10 nm Fe.sub.xO.sub.y Nanoparticles in Dimethyl Sulfoxide

(218) 5 nmol Fe.sub.xO.sub.y nanoparticles in toluene were precipitated by the addition of an equal volume of methanol. The cloudy mixture was centrifuged (6000g; 10 min) and the supernatant was discarded. The remaining pellet was dissolved in 1000 L chloroform and mixed with 6.9 mol (1390 fold excess) PI-b-PEO. The resulting solution was quickly injected into 9000 L DMSO and stirred for a minute. The clear, brownish colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 52).

EXAMPLE 31

(219) Composition of Micelle Encapsulated Magnetic, Oleic Acid, 10 nm Fe.sub.xO.sub.y Nanoparticles in N,N-dimethylformamide (DMF)

(220) 5 nmol Fe.sub.xO.sub.y nanoparticles in toluene were precipitated by the addition of an equal volume of methanol. The cloudy mixture was centrifuged (6000g; 10 min) and the supernatant was discarded. The remaining pellet was dissolved in 500 L chloroform and mixed with 6.9 mol (1390 fold excess) PI-b-PEO. The resulting solution was quickly injected into 9000 L N,N-dimethylformamide and stirred for a minute. The clear, brownish colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 53).

EXAMPLE 32

(221) Composition of Micelle Encapsulated Magnetic, Oleic Acid, 10 nm Fe.sub.xO.sub.y Nanoparticles in [BMIM][BF.sub.4]

(222) 5 nmol Fe.sub.xO.sub.y nanoparticles in toluene were precipitated by the addition of an equal volume of methanol. The cloudy mixture was centrifuged (6000g; 10 min) and the supernatant was discarded. The remaining pellet was dissolved in 500 L chloroform and mixed with 6.9 mol (1390 fold excess) PI-b-PEO. 100 L of the resulting solution were quickly injected into 1500 L [BMIM][BF.sub.4], were vigorously stirred and heated carefully to evaporate the remaining CHCl.sub.3. The clear, brownish colored solution thus obtained indicated the colloidal dissolution of the nanoparticles in polymer micelles. Micelle formation was confirmed by dynamic light scattering (DLS; FIG. 54).

EXAMPLE 33

(223) Composition of Micelle Encapsulated Fluorescent, Oleic Acid, 20 nm NaYF.sub.4:Yb/NaYF.sub.4 Nanoparticles in Methanol

(224) 9 nmol of NaYF4:Yb/NaYF4 nanoparticles in hexane were mixed with 30 mol (3333 fold excess) PI-DETA (2000 g/mol) and the volume was adjusted to 1 mL. After 2 h of incubation the nanoparticles were precipitated by the addition of 1200 L ethanol and centrifuged (10000g; 10 min). The supernatant was discarded and the pellet was dissolved in 900 L dichloromethane.

(225) 700 L (approx. 7 nmol) of this solution were mixed with 11.7 mol (1666 fold excess) PI-b-PEO and the total volume was adjusted to 2000 L. The resulting solution was quickly injected into 20 mL of methanol and stirred for a minute. The colorless solution was concentrated using a rotary evaporator (40 C.; 400 mbar) until a very concentrated solution (1-3 mL) was obtained. Subsequently, the volume was adjusted to 6 mL with methanol and divided into three equal parts. One of these parts was further diluted with 3 mL methanol (approx. 2.3 nmol NaYF.sub.4:Yb/NaYF.sub.4 nanoparticles) and characterized together with the native NaYF.sub.4:Yb/NaYF.sub.4 nanoparticles (2.3 nmol in 5 mL n-hexane). The encapsulation of the nanoparticles in polymeric micelles was concluded from the fact that no precipitation occurred after the injection into methanol, which is an excellent precipitant for the PI-DETA coated nanoparticles. Micelle formation was further confirmed by dynamic light scattering (DLS; FIG. 55). The emission properties of the nanoparticles are only slightly influenced by the encapsulation process (FIG. 56).