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Synthesis of highly luminescent colloidal particlesRelated Patent Categories: Stock Material Or Miscellaneous Articles, Coated Or Structually Defined Flake, Particle, Cell, Strand, Strand Portion, Rod, Filament, Macroscopic Fiber Or Mass Thereof, Particulate Matter (e.g., Sphere, Flake, Etc.), CoatedSynthesis of highly luminescent colloidal particles description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20060172133, Synthesis of highly luminescent colloidal particles. Brief Patent Description - Full Patent Description - Patent Application Claims
CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of and priority to U.S. Provisional Application Ser. No. 60/602,271 filed Aug. 17, 2004 the contents of which are incorporated herein by reference in their entirety. BACKGROUND [0002] Fluorescence-based analyses and nonisotopic detection systems have become a powerful tool for scientific research and clinical diagnostics for the detection of biomolecules using various assays including, but not limited to, flow cytometry, nucleic acid hybridization, DNA sequencing, nucleic acid amplification, immunoassays, histochemistry, and functional assays involving living cells. Fluorescent semiconductor nanocrystals have found widespread use due to their high fluorescent intensity and the ability of different nanocrystals to be excited by a single light source. It would be desirable to increase the signal from these non-isotopic materials to increase the sensitivity of a variety of assays and analyses utilizing them. It would be advantageous to controllably link numbers of nanocrystals and other small particles into structures for analytical applications that can be used to label a target molecule to be detected. [0003] Mirkin et al, in WO 98/04740 discloses nanoparticles having oligonucleotide attached to them. Methods are disclosed that comprise contacting a nucleic acid with one or more types of nanoparticles having oligonucleotides attached to them. The oligonucleotides are attached to nanoparticles and have sequences complementary to portions of the sequence of the nucleic acid. A detectable change, a color change, is brought about as a result of the hybridization of the oligonucleotides on the nanoparticles to the nucleic acid. The compositions disclosed do not include core/shell semiconductor nanocrystals and use oligonucleotides, specifically complementary oligonucleotides, to form conjugates of the nanoparticles. [0004] Mirkin et al. in U.S. Pat. No. 6,361,944 disclose nanoparticles having oligonucleotides attached to them and uses for the compositions. Again, the disclosure provides oligonucleotides attached to nanoparticles that include core/shell semiconductor nanocrystals and where the oligonucleotide sequences are complementary to portions of the sequence of a nucleic acid to be detected. A detectable change is brought about as a result of the hybridization of the oligonucleotides on the nanoparticles to the nucleic acid. The disclosure purports to illustrate the formation of nanoparticle aggregates, nanomaterials, and nanostructure by combining nanoparticles having complementary oligonucleotides attached to them, the nanoparticles being held together in the aggregates as a result of the hybridization of the complementary oligonucleotides. [0005] Hansen et al. in WO 98/33070 disclose a homogeneous binding assay. The disclosure describes a homogeneous method of measuring chemical binding that relies on resonant, or "amplified", optical extinction (light scattering plus absorption) from a defined, specific class of colloidal particles where the real term n of the complex refractive index n-ik approaches zero while the imaginary term k approaches 2.sup.(1/2). Chemical binding partners are coated onto the particles, which either aggregate or disperse during the binding reaction, causing an optical extinction change at one wavelength that is quantitatively related to the number of single colloidal particles and another at a second wavelength that is quantitatively related to the number of doublet colloidal particles. The disclosure describes the uses of optical extinction to measure the formation of particle dimers (through the appearance of increased extinction at the split resonant wavelength) and the concomitant disappearance of the singlet particles (through the decrease of extinction at the original resonant wavelength). [0006] Bawendi et al. in EP0990903 disclose biological applications of semiconductor nanocrystals. The disclosure describes compositions comprising fluorescent semiconductor nanocrystals associated to a compound, where the nanocrystals have a characteristic spectral emission that is tunable to a desired wavelength by controlling the size of the nanocrystal, and where the emission provides information about a biological state or event. [0007] Barbera-Guillem et al in U.S. Pat. No. 6,261,779 discloses nanocrystals having polynucleotide strands and their use to form dendrimers in a signal amplification system. The disclosure provides compositions and assay kits comprising functionalized nanocrystals having a plurality of polynucleotide strands of known sequence extending from them. The disclosure describes primary dots that are used to operably link to a molecule, and secondary dots comprise a plurality of polynucleotide strands which are complemetary to the plurality of polynucleotide strands of the primary dots. The disclosure provides a method for detecting the presence or absence of target molecules in a sample comprising operably linking primary dots to molecules, contacting the complex formed with the sample, contacting the sample with successive additions of secondary dots and primary dots. If a target molecule is present in the sample, the primary dots and secondary dots will form dendrimers that can be detected by fluorescence emission. [0008] Peng et al. U.S. Pat. No. 6,872,249 disclose the synthesis of colloidal nanocrystals. A method of synthesizing colloidal nanocrystals is disclosed using metal oxides or metal salts as a precursor. The metal oxides or metal salts are combined with a ligand and then heated in combination with a coordinating solvent. [0009] Peng et al. U.S. Pat. No. 6,869,545 discloses colloidal nanocrystals with high photoluminescence quantum yields and methods of preparing the same. The disclosure provides compositions containing colloidal nanocrystals with high photoluminescence quantum yields, synthetic methods for the preparation of highly luminescent colloidal nanocrystals, as well as methods to control the photoluminescent properties of colloidal nanocrystals. [0010] Bawendi et al. in U.S. Pat. No. 6,306,610 disclose quantum dot white and colored light emitting diodes. The disclosure describes an electronic device comprising a population of quantum dots embedded in a host matrix and a primary light source which causes the dots to emit secondary light of a selected color, and a method of making such a device. The size distribution of the quantum dots is chosen to allow light of a particular color to be emitted from the structure. The dots can be composed of an undoped semiconductor such as CdSe, and may optionally be overcoated to increase photoluminescence. The host matrix for the device includes isolated dots within the matrix and not defined aggregates of nanocrystals. [0011] U.S. Pub. No. 20040110220 to Mirkin et al. discloses nanoparticles having oligonucleotides attached to them and uses for such coated nanoparticles. The disclosure provides methods of detecting a nucleic acid that comprise contacting the nucleic acid with one or more types of nanoparticles having oligonucleotides attached to them. The disclosure describes a method where oligonucleotides are attached to nanoparticles and have sequences complementary to portions of the sequence of the nucleic acid. A detectable change is brought about as a result of the hybridization of the oligonucleotides on the nanoparticles to the nucleic acid. The disclosure describes methods of synthesizing nanoparticle-oligonucleotide conjugates and methods of using the conjugates. The disclosure describes nanomaterials and nanostructures comprising nanoparticles and methods of nanofabrication utilizing nanoparticles. The disclosure describes a method of separating a selected nucleic acid from other nucleic acids. SUMMARY [0012] There is a need to form colloidal aggregates or cluster of particles in a controlled manner using inexpensive coating materials. Such clusters could be used in a variety of nonisotopic detection systems to increase the signal comprising fluorescence emission of high quantum yield. Such clusters may be used to provide tailored signal amplification that is not limited as to the chemical nature of the target molecule to be detected. It would be desirable that such non-isotopic probes could be used to bind target molecules of various and that they can be excited with a single excitation light source and with resultant fluorescence emissions with discrete fluorescence peaks. [0013] Embodiments of the invention include nanocrystal aggregates. These compositions can include two or more aggregated nanocrystals; where the nanocrystals includes a coating layer, and the coating layer can include one or more imidazole groups. The coated nanocrystals interact or associate through their coating layers to form an aggregate. [0014] The composition of aggregated nanocrystals may further include a cross linking agent. The aggregated nanocrystals can be crosslinked by one or more organophosphine compounds. The cross linking agent can include tris(hydroxy methyl) phosphine, beta-[tris(hydroxymethyl)phosphino]propionic acid, any combination of these, or other suitable organophosphine compounds. [0015] The coating layer on the nanocrystals can be bound or operably linked to the nanocrystal by the one or more imidazole groups. The coating layer on the nanocrystals can include histidine, carnosine, polyhistidine, polyimidazole, glycyl histidine or other similar imidizole containing compounds. In some embodiments one or more imidazole groups of the coating layer bond or otherwise operably link the imidazole coating compound to the nanocrystal. [0016] The aggregated nanocrystals can be luminescent, fluorescent, magnetic, or may include one or more these properties by aggregating two or more different nanocrystals that include any of these properties. In some embodiments, the aggregates can include nanocrystals that are semiconductor core nanocrystals or semiconductor core/shell nanocrystals. [0017] The aggregate may further comprise at least one functional group on the surface of the aggregate. The aggregate may further comprise functional groups on the surface of the coated nanocrystal aggregate such as but not limited to hydroxyl, thiol, amino, acetylenic, carboxyl, ester, amide, dicarboxylic, carboxamide selenol, hydrazide, aldehyde, or combinations of any of these. The aggregates can be dispersed in a variety of organic solvents, mixtures of organic solvent and water, or in aqueous based solutions. [0018] Embodiments of the invention can include aggregates that have been functionalized with reactive groups. A functionalized aggregate composition can include a nanocrystal aggregate and at least one affinity molecule. The nanocrystal aggregate comprises two or more coated nanocrystals; the nanocrystals comprise a coating layer comprising one or more imidazole groups where the nanocrystals interact through their coating layers to form an aggregate. In some embodiments, the aggregate may include at least one functional group on its surface with at least one affinity molecule is linked to the functional group. [0019] In embodiments of the invention the affinity molecule can be but is not limited to a polyclonal antibody, a monoclonal antibody, a peptide, an aptamer, a nucleic acid, a polynucleotide, a lectin, a lipid, a small organic molecule, a polysaccharide, avidin, neutravidin, streptavidin, an avidin derivative, biotin, a biotin derivative, or any combination of these affinity molecules. The affinity molecule can be covalently linked to the functional group. The functional group can include but is not limited to hydroxyl, thiol, amino, carboxyl, ester, amide, dicarboxylic, carboxamide, selenol, hydrazide, aldehyde, or any combination of these. [0020] Some embodiments of the invention can include an aggregate having a defined number of coated nanocrystals, coated nanoparticles, or any combination of these in the aggregate. The aggregate composition can include from about two or more to about twenty aggregated coated fluorescent semiconductor nanocrystals. The coated nanocrystals and or coated nanoparticles in the aggregate comprise a coating layer that includes one or more imidazole groups and the coated nanocrystals interact through their coating to form the aggregate. In some embodiments one or more imidazole groups of the coating layer bond or otherwise operably link the imidazole coating compound to the nanocrystals or nanoparticles. The number of coated nanocrystals, coated nanoparticles, or combination of these in the aggregate or cluster can separately form a cluster of a defined size, preferably between 2 and 20 coated nanocrystals of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 coated nanometer sized particles. The cluster or aggregate is a colloidal particle and the nanocrystals and/or nanoparticles in the aggregate can be crosslinked by an organophosphine compound. [0021] One embodiment of the invention is a method of preparing aggregates comprised of coated nanocrystals. A method of preparing these aggregates can include the acts or steps of providing two or more nanocrystals, the nanocrystals include a coating layer comprising at least one imidazole group, and then contacting or combining the nanocrystals to prepare an aggregate coated nanocrystals. The nanocrystals interact through their coating layers to form the aggregate. The method can further include the act of placing the two or more nanocrystals in a solvent mixture and contacting or combining them to form an aggregate or an aggregate of a predetermined size. The solvent mixture can be an aqueous solvent mixture. Similar acts or steps can be used to prepare clusters or aggregates that include coated nanoparticles, or any combination of coated nanoparticles and coated nanocrystals where the coating layer comprises at least one imidazole group. Continue reading about Synthesis of highly luminescent colloidal particles... 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