Methods for forming dyed microspheres and populations of dyed microspheres

• US 20060085925A1
• Filed: 10/11/2005
• Published: 04/27/2006
• Est. Priority Date: 10/12/2004
• Status: Active Grant

• Current Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Original Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Inventors: Lugade, Ananda G., Hoffacker, Kurt D., Terpetschnig, Ewald
• Granted Patent: US 8,038,734 B2
• Time in Patent Office: Days
• US Class Current: 8/485
• CPC Class Codes: C08F 212/08 Styrene
• CPC Class Codes: C08F 8/00 Chemical modification by after-treatment graft polymers, block polymers, crosslinking with unsaturated monomers or with polymers C08F251/00 - C08F299/00; of conjugated diene rubbers C08C
• CPC Class Codes: C09B 67/0097 Dye preparations of special physical nature; Tablets, films, extrusion, microcapsules, sheets, pads, bags with dyes
• CPC Class Codes: C09B 69/10 Polymeric dyes; Reaction products of dyes with monomers or with macromolecular compounds addition products of alkylene oxide to dyes, C09B69/00; dyeing with polymeric dyes D06P1/0056
• CPC Class Codes: D06P 1/008 Preparing dyes in situ D06P1/12, D06P1/145 take precedence
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex

Abstract

Various methods for forming dyed microspheres are provided. One method includes activating a chemical structure coupled to a dye using heat or light to form a reaction intermediate in the presence of a microsphere. The reaction intermediate covalently attaches to a polymer of the microsphere thereby coupling the dye to the polymer and forming the dyed microsphere. Additional methods are provided for forming a dyed microsphere coupled to a molecule. These methods include dyeing the microspheres as described above in addition to synthesizing the molecule on an outer surface of the dyed microspheres. A population of dyed microspheres is also provided. Each of the dyed microspheres of the population includes a dye attached to a polymer of each of the dyed microspheres by a chemical structure. A coefficient of variation in dye characteristics of the population of dyed microspheres attributable to the dye is less than about 10%.

Claims

1. A method for forming a dyed microsphere, comprising activating a chemical structure coupled to a dye using heat or light to form a reaction intermediate in the presence of a microsphere, wherein the reaction intermediate covalently attaches to a polymer of the microsphere thereby coupling the dye to the polymer and forming the dyed microsphere.

2. The method of claim 1, wherein said activating comprises heating the chemical structure coupled to the dye to a temperature of about 110°

C.

3. The method of claim 1, wherein said activating comprises heating the chemical structure coupled to the dye to a temperature of about 50°

C. to about 80°

C.

4. The method of claim 1, wherein said activating comprises heating the chemical structure coupled to the dye to a temperature below the glass transition temperature of the polymer.

5. The method of claim 1, wherein the microsphere has substantially the same shape before and after said activating.

6. The method of claim 1, wherein the dye has substantially the same structure and substantially the same composition before and after said activating.

7. The method of claim 1, wherein the chemical structure and the dye do not detach from the polymer when the dyed microsphere is disposed in an organic solvent.

8. The method of claim 1, wherein the polymer consists essentially of polystyrene crosslinked with divinyl benzene.

9. The method of claim 1, wherein the dyed microsphere comprises one or more functional groups attached to the polymer such that one or more molecules can be attached to the dyed microsphere via the one or more functional groups.

10. The method of claim 1, wherein the chemical structure comprises an aromatic sulfonyl azide.

11. The method of claim 1, wherein the chemical structure comprises 1,5-ethyl-di-amino-naphthalene-sulfonic azide.

12. The method of claim 1, wherein the reaction intermediate comprises a sulfonyl nitrene group.

13. The method of claim 1, wherein the reaction intermediate comprises a carbene group or a benzophenone triplet.

14. The method of claim 1, wherein the reaction intermediate comprises a functional group separated from the dye by a non-activated portion of the chemical structure.

15. The method of claim 1, wherein the reaction intermediate comprises a functional group located on one side of a non-activated portion of the chemical structure, and wherein the dye is located on another side of the non-activated portion substantially opposite to the one side.

16. The method of claim 1, further comprising activating an additional chemical structure coupled to an additional, different dye using heat or light to form a different reaction intermediate in the presence of the microsphere, wherein the different reaction intermediate covalently attaches to the polymer of the microsphere thereby coupling the additional, different dye to the polymer.

17. The method of claim 16, wherein the additional, different dye has substantially the same structure and composition before and after said activating.

18. A method for forming a dyed microsphere coupled to a molecule, comprising:

activating a chemical structure coupled to a dye using heat or light to form a reaction intermediate in the presence of a microsphere, wherein the reaction intermediate covalently attaches to a polymer of the microsphere thereby coupling the dye to the polymer and forming the dyed microsphere; and

synthesizing the molecule on an outer surface of the dyed microsphere.

19. The method of claim 18, wherein said synthesizing comprises disposing the dyed microsphere in an organic solvent.

20. The method of claim 18, wherein said synthesizing comprises disposing the dyed microsphere in an organic solvent, and wherein the dyed microsphere has substantially the same dye characteristics before and after said synthesizing.

21. The method of claim 18, wherein the molecule comprises a biomolecule.

22. The method of claim 18, wherein the molecule comprises an oligonucleotide, a peptide, or an oligosaccharide.

23. The method of claim 18, wherein the molecule comprises an organic molecule.

24. The method of claim 18, wherein the molecule is substantially insoluble in aqueous solvents.

25. The method of claim 18, wherein said synthesizing comprises disposing the dyed microsphere in an aqueous solvent.

26. The method of claim 18, wherein said activating comprises heating the chemical structure coupled to the dye to a temperature of about 110°

C.

27. The method of claim 18, wherein said activating comprises heating the chemical structure coupled to the dye to a temperature of about 50°

C. to about 80°

C.

28. The method of claim 18, wherein said activating comprises heating the chemical structure coupled to the dye to a temperature below the glass transition temperature of the polymer.

29. The method of claim 18, wherein the microsphere has substantially the same shape before and after said activating.

30. The method of claim 18, wherein the dye has substantially the same structure and substantially the same composition before and after said activating.

31. The method of claim 18, wherein the chemical structure and the dye do not detach from the polymer when the dyed microsphere is disposed in an organic solvent.

32. The method of claim 18, wherein the polymer consists essentially of polystyrene crosslinked with divinyl benzene.

33. The method of claim 18, wherein the dyed microsphere comprises one or more functional groups attached to the polymer such that the molecule can be synthesized on the outer surface of the dyed microsphere via the one or more functional groups.

34. The method of claim 18, wherein the chemical structure comprises an aromatic sulfonyl azide.

35. The method of claim 18, wherein the chemical structure comprises 1,5-ethyl-di-amino-naphthalene-sulfonic azide.

36. The method of claim 18, wherein the reaction intermediate comprises a sulfonyl nitrene group.

37. The method of claim 18, wherein the reaction intermediate comprises a carbene group or a benzophenone triplet.

38. The method of claim 18, wherein the reaction intermediate comprises a functional group separated from the dye by a non-activated portion of the chemical structure.

39. The method of claim 18, wherein the reaction intermediate comprises a functional group located on one side of a non-activated portion of the chemical structure, and wherein the dye is located on another side of the non-activated portion substantially opposite to the one side.

40. The method of claim 18, further comprising activating an additional chemical structure coupled to an additional, different dye using heat or light to form a different reaction intermediate in the presence of the microsphere, wherein the different reaction intermediate covalently attaches to the polymer of the microsphere thereby coupling the additional, different dye to the polymer.

41. The method of claim 40, wherein the additional, different dye has substantially the same structure and composition before and after said activating.

42. A population of dyed microspheres, wherein each of the dyed microspheres of the population comprises a dye attached to a polymer of each of the dyed microspheres by a chemical structure, and wherein a coefficient of variation in dye characteristics of the population of dyed microspheres attributable to the dye is less than about 10%.

43. The population of claim 42, wherein the coefficient of variation is a coefficient of variation in a signal emitted by each of the dyed microspheres upon excitation of each of the dyed microspheres.

44. The population of claim 42, wherein an amount of the dye attached to the polymer of each of the dyed microspheres is substantially equal.

45. The population of claim 42, wherein each of the dyed microspheres of the population further comprises an additional, different dye attached to the polymer of each of the dyed microspheres by an additional chemical structure.

46. The population of claim 45, wherein the coefficient of variation in the dye characteristics of the population of dyed microspheres attributable to the additional, different dye is less than about 10%.

47. The population of claim 42, wherein each of the dyed microspheres of the population further comprises a biomolecule synthesized on an outer surface of each of the dyed microspheres.

48. The population of claim 42, wherein each of the dyed microspheres of the population further comprises an organic molecule synthesized on an outer surface of each of the dyed microspheres.

49. The population of claim 42, wherein each of the dyed microspheres has substantially the same shape before and after the dye is attached to the polymer.

50. The population of claim 42, wherein the dye has substantially the same structure and substantially the same composition before and after the dye is attached to the polymer.

51. The population of claim 42, wherein the chemical structure and the dye do not detach from the polymer when the population of dyed microspheres is disposed in an organic solvent.

52. The population of claim 42, wherein the polymer consists essentially of polystyrene crosslinked with divinyl benzene.

53. The population of claim 42, wherein each of the dyed microspheres further comprises one or more functional groups attached to the polymer of each of the dyed microspheres such that one or more molecules can be attached to each of the dyed microspheres via the one or more functional groups.

54. The population of claim 42, wherein the chemical structure comprises an aromatic sulfonyl azide.

55. The population of claim 42, wherein the chemical structure comprises 1,5-ethyl-di-amino-naphthalene-sulfonic azide.