Functionalized fluorescent nanocrystal compositions and methods of making

• US 7,205,048 B2
• Filed: 04/09/2003
• Issued: 04/17/2007
• Est. Priority Date: 09/17/2001
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Invitrogen Corp. (Thermo Fisher Scientific Incorporated)
• Inventors: Naasani, Imad
• Primary Examiner(s): KILIMAN, LESZEK B
• Application Number:
  US10/409,940
• Publication Number:
  US 20040247861A1
• Time in Patent Office: 1,469 Days
• Field of Search: 428/403, 428/407, 428/690, 252/301.4 R, 252/301.4 S, 252/301.6 R, 252/301.6 S
• US Class Current: 428/403
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: G01N 33/588 with semiconductor nanocrystal label, e.g. quantum dots
• CPC Class Codes: Y10T 428/265 1 mil or less
• CPC Class Codes: Y10T 428/2982 Particulate matter [e.g., sphere, flake, etc.]
• CPC Class Codes: Y10T 428/2989 Microcapsule with solid core [includes liposome]
• CPC Class Codes: Y10T 428/2991 Coated
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer

Abstract

The present invention provides for functionalized fluorescent nanocrystal compositions and methods for making these compositions. The compositions are fluorescent nanocrystals coated with at least one material. The coating material has chemical compounds or ligands with functional groups or moieties with conjugated electrons and moieties for imparting solubility to coated fluorescent nanocrystals in aqueous solutions. The coating material provides for functionalized fluorescent nanocrystal compositions which are water soluble, chemically stable, and emit light with a high quantum yield and/or luminescence efficiency when excited with light. The coating material may also have chemical compounds or ligands with moieties for bonding to target molecules and cells as well as moieties for cross-linking the coating. In the presence of reagents suitable for reacting to form capping layers, the compounds in the coating may form a capping layer on the fluorescent nanocrystal with the coating compounds operably bonded to the capping layer.

Claims

1. A functionalized fluorescent core/shell nanocrystal comprised of:

a fluorescent core/shell nanocrystal;

a coating material, said coating material being histidine-based.

2. The functionalized fluorescent nanocrystal of claim 1, wherein said fluorescent nanocrystal emits light with a quantum yield of greater than about 10%.

3. The functionalized fluorescent nanocrystal of claim 1, wherein said fluorescent nanocrystal emits light with a quantum yield of greater than about 30%.

4. The functionalized fluorescent nanocrystal of claim 1, wherein said fluorescent nanocrystal emits light with a quantum yield of greater than about 50%.

5. The functionalized fluorescent nanocrystal of claim 1, wherein said fluorescent nanocrystal emits light with a quantum yield of greater than about 70%.

6. The functionalized fluorescent nanocrystal of claim 1, wherein said coating material includes carnosine.

7. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal is soluble in a fluid with a surface energy of greater than 35 dynes/cm.

8. The functionalized fluorescent nanocrystal of claim 1, wherein said fluorescent nanocrystals are soluble in an aqueous solution.

9. The functionalized fluorescent nanocrystal of claim 1, further comprising a molecular probe operably bound to said coating material, wherein said molecular probe includes a polynucleotide of known sequence attached to said nanocrystal via an amino terminal portion of said polynucleotide.

10. The functionalized fluorescent nanocrystal of claim 1, further comprising a molecular probe operably bound to said coating material, wherein said molecular probe includes a polynucleotide of known sequence attached to said nanocrystal via a carboxy terminal portion of said polynucleotide.

11. The functionalized fluorescent nanocrystal composition of claim 1, wherein said fluorescent nanocrystal is cadmium selenide;

and said coating material comprises zinc sulfide operably bonded to cross linked histidine and (beta-(tris(hydroxymethyl)phosphino)propionic acid)).

12. The functionalized fluorescent nanocrystal composition of claim 1, wherein said fluorescent nanocrystal is cadmium selenide;

and said coating material comprises zinc sulfide operably bonded to cross linked carnosine and (beta-(tris(hydroxymethyl)phosphino)propionic acid)).

13. The functionalized fluorescent nanocrystal composition of claim 1, further comprising a molecular probe operably bound to said coating material wherein said molecular probe is chosen from the group consisting of avidin, ConA, lectin, and IgG.

14. The functionalized fluorescent nanocrystal composition of claim 13, wherein said molecular probe is avidin.

15. The functionalized fluorescent nanocrystal of claim 1, wherein said nanocrystal is a doped metal oxide nanocrystal.

16. The functionalized fluorescent nanocrystal of claim 15, wherein said doped metal oxide nanocrystal is magnetized when irradiated with light.

17. The functionalized fluorescent nanocrystal of claim 1, wherein said fluorescent nanocrystals have a characteristic spectral emission intensity which is enhanced by the coating material.

18. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal comprises a core material and a shell material.

19. The functionalized fluorescent nanocrystal of claim 18, wherein said core material is CdSe and said shell material is ZnS.

20. The functionalized fluorescent nanocrystal of claim 17, wherein said emission provides information about a biological state or event.

21. A method for detecting a target species, the method comprising:

exciting a complex formed by operably bonding a fluorescent core/shell nanocrystal to a target molecule to form a complex wherein said fluorescent nanocrystal includes a histidine-based coating material; and

detecting the light emitted from said complex.

22. The method of claim 21, wherein the detecting further comprises detecting the intensity of light emitted from said complex in the perpendicular and parallel directions.

23. A method for separating materials comprising:

introducing doped metal oxide fluorescent core/shell nanocrystal soluble in fluids being coated with the histidine-based coating material;

exciting said fluorescent nanocrystal to have a magnetic moment; and

utilizing a magnetic field for separating the doped metal oxide fluorescent nanocrystals with the induced magnetic moment.

24. The method of claim 23, wherein said doped metal oxide fluorescent nanocrystals further comprise a molecule operably bound to said doped metal oxide fluorescent nanocrystal.

25. The method of claim 24, wherein said molecule is chosen from the group consisting of target molecules and molecular probes.

26. A functionalized fluorescent nanocrystal composition comprising a fluorescent core/shell nanocrystal, a histidine-based coating material, and a molecular probe wherein said fluorescent nanocrystals have a characteristic spectral emission which is enhanced by the coating material.

27. The composition of claim 26, wherein the coating material is a carnosine-based composition.

28. The composition of claim 26, wherein the fluorescent nanocrystal comprises a core material and a shell material.

29. The composition of claim 28, wherein said core material is CdSe and said shell material is ZnS.

30. The composition of claim 26, wherein said emission provides information about a biological state or event.

31. A coated nanocrystal comprising:

a core comprising a first semiconductor material, a shell layer deposited on the core comprising a second semiconductor material, a coating material deposited on said shell layer including a histidine-based composition coating material wherein said coated nanocrystal has a quantum yield of greater than 50% when irradiated with light.

32. The coated nanocrystal of claim 31, wherein the second semiconductor material is ZnS.

33. The coated nanocrystal of claim 31, wherein the shell layer is from about one to about two monolayers of the second semiconductor material.

34. The coated nanocrystal of claim 31, wherein the second semiconductor material is ZnSe.

35. The coated nanocrystal of claim 31, wherein the nanocrystal further comprises an organic layer on the nanocrystal coating layer.

36. The coated nanocrystal of claim 35, wherein the organic layer comprises a moiety selected to provide compatibility with a suspension medium.

37. The coated nanocrystal of claim 35, wherein the organic layer comprises a moiety selected to exhibit affinity for the outer surface of the nanocrystal.

38. The coated nanocrystal of claim 37, wherein the organic layer comprises a short-chain polymer terminating in a moiety having affinity for a suspending medium.

39. The coated nanocrystal of claim 31, wherein the first semiconductor material is selected from the group consisting of CdS, CdSe, CdTe, and mixtures thereof.

40. The coated nanocrystal of claim 39, wherein the second semiconductor material has a higher band gap than the first semiconductor material.

41. The coated nanocrystal of claim 31, wherein the second semiconductor material is selected from the group consisting of ZnS, ZnSe, CdS, CdSe, and mixtures thereof.

42. The coated nanocrystal of claim 31, wherein the first semiconductor material is CdSe and the second semiconductor material is ZnS.

43. The coated nanocrystal of claim 42, wherein the shell layer comprises greater than about 0 to about 5.3 monolayers of the second semiconductor material.

44. The coated nanocrystal of claim 43, wherein the shell layer comprises less than about one monolayer of the second semiconductor material.

45. The coated nanocrystal of claim 44, wherein the second semiconductor material is ZnS or ZnSe.

46. The coated nanocrystal of claim 43, wherein the shell layer comprises in the range of about one to about two monolayers of the second semiconductor material.

47. The coated nanocrystal of claim 43, wherein the second semiconductor material is ZnS or ZnSe.

48. The coated nanocrystal of claim 31, wherein said coated nanocrystal is a member of a substantially monodisperse particle population.