초록 ▼

A dye, such as a fluorescent dye, is incorporated into polymer microparticles using a solvent system composed of a first solvent in which the dye and the microparticle polymer are soluble, a second solvent in which the dye and the microparticle polymer are not or only weakly soluble, and a third sol

A dye, such as a fluorescent dye, is incorporated into polymer microparticles using a solvent system composed of a first solvent in which the dye and the microparticle polymer are soluble, a second solvent in which the dye and the microparticle polymer are not or only weakly soluble, and a third solvent in which the dye and the microparticle polymer are not or only weakly soluble. The first and second solvents are immiscible with each other, or at most partially miscible. The third solvent is miscible with the first and second solvents. The formulation provides substantially complete partitioning of the dye to the microparticles. The method may be used to obtain dyed polymer microparticle formed of cross-linked or non-cross-linked polymers. Libraries are provided comprising two or more sets of microparticles of different dye loadings. Fluorescent core-shell microparticles are produced from a mixture of microparticle cores incorporating one or more fluorescent dyes, a polymerization mixture comprising at least one polymerizable shell monomer, at least one free radical polymerization initiator comprising a water-insoluble oxidizing agent, and at least one water-soluble reducing agent.

대표청구항 ▼

1. A method of incorporating at least one dye into polymer microparticles comprising:(a) providing: (i) at least one first solvent in which the dye and the microparticle polymer are soluble; (ii) at least one second solvent in which the dye and the microparticle polymer are not or only weakly solubl

1. A method of incorporating at least one dye into polymer microparticles comprising:(a) providing: (i) at least one first solvent in which the dye and the microparticle polymer are soluble; (ii) at least one second solvent in which the dye and the microparticle polymer are not or only weakly soluble, said first and second solvents being immiscible or at most partially miscible; (iii) at least one third solvent in which the dye and the microparticle polymer are not or only weakly soluble, said third solvent being miscible with the first and second solvents; (b) suspending the polymer microparticles in a designated volume of a mixture comprising at least one second solvent and at least one third solvent; (c) adding to said polymer microparticle suspension a dye solution comprising at least one first solvent and at least one dye dissolved therein, the amount of dye corresponding to the desired final state of dye incorporation in the microparticles; (d) adding to said polymer microparticle suspension at least one second solvent to induce substantially complete partitioning of the dye from the suspension liquid phase to the microparticles. 2. The method according to claim 1 wherein the rate of addition of the second solvent to the polymer microparticle suspension in step (d) is controlled to maintain phase stability in the suspension.3. The method according to claim 1 wherein the mixture comprising the least one second solvent and the at least one third solvent further comprises at least one suspension stabilizer.4. The method according to claim 1 wherein the dye is a fluorescent dye.5. The method according to claim 4 wherein the dye is a hydrophobic dye.6. The method according to claim 5 wherein the dye is selected from the group consisting of styryl dyes, pyrromethane dyes, coumarin dyes, and combinations thereof.7. The method according to claim 1 wherein the microparticles comprise a hydrophobic polymer.8. The method according to claim 7 wherein the polymer is a homopolymer or copolymer comprising a vinyl-containing monomer.9. The method according to claim 7 selected from the group consisting of homopolymers or copolymers of polystyrene, poly(methyl methacrylate), polyacrylamide, poly(ethylene glycol), poly(hydroxyethylmethacrylate), poly(vinyltoluene), poly(divinylbenzene), and combinations thereof.10. The method according to claim 8 wherein the polymer is polystyrene or copolymer thereof containing at least 50% by weight styrene monomer units.11. The method according to claim 10 wherein the polymer is a styrene/methacrylic acid copolymer.12. The method according to claim 8 wherein the polymer is non-cross-linked.13. The method according to claim 8 wherein the microparticles have a diameter of from about 0.1 to about 100 microns.14. The method according to claim 1 wherein the particles are monodisperse.15. The method according to claim 1 wherein the concentration of dye present in the microparticle suspension formed by contacting microparticles with said dye solution is from about 10 μg/g to about 100 μg/g, based upon the weight of the microparticle suspension.16. The method according to claim 15 comprising a solvent wherein the first solvent is selected from the group consisting of methylene chloride, chloroform, tetrahydrofuran, dioxane, cyclohexane, benzene, toluene, butylacetate, lower chlorinated aliphatic hydrocarbons, and combinations thereof; the second solvent is water; and the third solvent is selected from the group consisting of acetone, lower alcohols, and combinations thereof.17. The method according to claim 16 wherein the first solvent is methylene chloride or dichloromethane.18. The method according to claim 17 wherein the third solvent is an alcohol.19. The method according to claim 1 wherein the microparticle is a core-shell microparticle comprising a central core comprising one or more core polymers surrounded by a shell comprising one or more shell polymers.20. The method according to claim 19 wherein the core polymer comprises a copolymers of styrene and a monomer more hydrophilic than styrene.21. The method according to claim 19 wherein the core polymer comprises methacrylic acid.22. The method according to claim 19 wherein the shell surface comprises one or more functional groups selected from the group consisting of sulfate, phosphate, hydroxyl, carboxyl, ester, amide, amidine, amine, sulfhydryl and aldehyde.23. The method according to claim 19 wherein the shell surface comprises an attached oligonucleotide, polynucleotide, peptide or polypeptide.24. The method according to claim 1 wherein at least a portion of the microparticles are magnetically responsive.25. A method for producing fluorescent core-shell microparticles comprising:(a) providing microparticle cores incorporating one or more fluorescent dyes; (b) contacting said microparticle cores with a polymerization mixture comprising at least one polymerizable shell monomer, at least one free radical polymerization initiator comprising a water-insoluble oxidizing agent, and at least one water-soluble reducing agent; (c) polymerizing said shell monomer to form a polymer shell around said microparticle core. 26. The method according to claim 25 wherein the polymerization mixture comprises an oil-in-water emulsion.27. The method according to claim 25 wherein contacting the microparticle cores with the polymerization mixture comprises the steps of swelling the cores with a first mixture comprising said shell monomer and water-insoluble oxidizing agent to provide uptake of said first mixture into the cores, and then contacting the swelled microparticle cores with the water-soluble reducing agent.28. The method according to claim 25 wherein the polymerization mixture further comprises at least one water-soluble free radical scavenger.29. The method according to claim 25 wherein the shell monomer is more hydrophilic that the polymer comprising the microparticle core.30. The method according to claim 29 wherein the microparticle core polymer comprises polystyrene or copolymer thereof and the shell monomer comprises methyl methacrylate.31. The method according to claim 30 wherein the shell monomer comprises a mixture of methyl methacrylate, hydroxyethyl methacrylate and methacrylic acid.32. The method according to claim 25 wherein the water-insoluble oxidizing agent is selected from the group consisting of hydrophobic organic peroxides and hydrophilic organic hydroperoxides.33. The method according to claim 25 wherein the water-insoluble oxidizing agent is selected from the group consisting of cumenhydroperoxide and t-butylperoxy-2-ethylhexanoate.34. The method according to claim 25 wherein the water-soluble reducing agent is selected from the group consisting of tetraethylenepentamine and sodium formaldehydesulfoxylate-Fe(II) ethylene diamine tetra acetic acid.35. The method according to claim 25 wherein the microparticle cores have been stained with fluorescent dye according to the method of claim 1.36. A method of preparing a library of dyed polymer microparticles comprising two or more sets of microparticles of different dye loading comprising:forming a first set of microparticles according to the method of claim 1, the microparticles of said first set characterized by a first dye loading; forming a second set of microparticles according to the method of claim 1, the microparticles of said second set characterized by a second dye loading different from said first dye loading. 37. The method according to claim 36 wherein the difference in dye loading between the microparticles of said first set and the microparticles of said second set comprises a difference in at least one of dye concentration and dye identity.38. The method according to claim 37 wherein the difference in dye loading between the microparticles of said first set and the microparticles of said second set comprises a difference in dye concentration.39. The method according to claim 37 wherein the difference in dye loading between the microparticles of said first set and the microparticles of said second set comprises a difference in dye identity.40. The method according to claim 36 wherein the microparticles comprise non-cross-linked polymers.41. The microparticle according to claim 40 wherein the dye is substantially uniformly distributed throughout said microparticle.42. The method according to claim 36 wherein the first and second dye loadings comprise one or more fluorescent dyes.43. The method according to claim 42 wherein the dyes are selected from the group consisting of styryl dyes, pyrromethane dyes, coumarin dyes, and combinations thereof.44. The method according to claim 36 wherein the first and second dye loadings comprise one or more hydrophobic dyes.45. The method according to claim 44 wherein the polymer is a homopolymer or copolymer comprising a vinyl-containing monomer.46. The method according to claim to claim 44 wherein the polymer is a styrene/methacrylic acid copolymer.