Highly luminescent functionalized semiconductor nanocrystals for biological and physical applications

• US 7,214,428 B2
• Filed: 04/09/2003
• Issued: 05/08/2007
• 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:
  US10/410,108
• Publication Number:
  US 20040009341A1
• Time in Patent Office: 1,490 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 428/25 Web or sheet containing structurally defined element or component and including a second component containing structurally defined particles
• CPC Class Codes: Y10T 428/2991 Coated
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer
• CPC Class Codes: Y10T 428/31721 Of polyimide

Abstract

The present invention provides for detection apparatus with functionalized fluorescent nanocrystal compositions and methods for making and using these compositions in biological detection applications, material separations, and in the production of biosensors. The compositions are fluorescent nanocrystals coated with at least one coating material comprising ligands with functional groups or moieties with conjugated electrons and moieties for imparting solubility to 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.

Claims

1. A functionalized fluorescent nanocrystal probe comprised of:

a fluorescent nanocrystal;

a coating material coating said fluorescent nanocrystal, said coating material being imidazole-based and further comprising a phosphine cross-linking compound; and

a molecular probe material operably bound to said coating material.

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

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

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

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

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

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

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

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

10. The functionalized fluorescent nanocrystal probe of claim 1, wherein said probe includes a polynucleotide of known sequence attached to said nanocrystal via a carboxy terminal portion of said polynucleotide.

11. The functionalized fluorescent nanocrystal probe of claim 1, wherein said fluorescent nanocrystal is cadmium selenide with a capping material including zinc sulfide that is operably bonded to cross linked histidine and (beta-(tris(hydroxymethyl)phosphino)propionic acid)).

12. The functionalized fluorescent nanocrystal probe 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(hydroxyrnethyl)phosphino)propionic acid)).

13. The functionalized fluorescent nanocrystal probe of claim 1, wherein said molecular probe is chosen from the group consisting of avidin, ConA, lectin, and IgG.

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

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

16. The functionalized fluorescent nanocrystal probe of claim 1, wherein said doped metal oxide nanocrystal is magnetized when irradiated with light.

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

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

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

20. The functionalized fluorescent nanocrystal probe 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 nanocrystal to a target molecule to form a complex wherein said fluorescent nanocrystal includes a imidazole-based coating material, wherein said coating material is further comprised of a phosphine cross-linking compound; 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 perpendicular and parallel to an excitation source.

23. A method for separating materials comprising:

introducing a doped metal oxide fluorescent core/shell nanocrystal soluble in fluids with a surface energy of at least 35 dynes/cm into a system;

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 connected 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 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 comprised of an imidazole-based composition wherein said coated nanocrystal has a quantum yield of greater than 30% when excited with light.

27. The coated nanocrystal of claim 26, wherein the nanocrystal has a quantum yield of greater than 50% when excited with light.

28. The coated nanocrystal of claim 26, wherein the second semiconductor material is ZnS.

29. The coated nanocrystal of claim 26, wherein the second semiconductor material is ZnSe.

30. The coated nanocrystal of claim 26, wherein the first semiconductor material is selected from the group consisting of CdS, CdSe, CdTe, and mixtures thereof.

31. The coated nanocrystal of claim 30, wherein the second semiconductor material has a higher band gap than the first semiconductor.

32. The coated nanocrystal of claim 30, wherein the second semiconductor material is selected from the group consisting of ZnS, ZnSe, CdS, CdSe, and mixtures thereof.

33. The coated nanocrystal of claim 30, wherein the first semiconductor material is CdSe and the second semiconductor material is ZnS.