Fluorescence energy transfer in particles
First Claim
1. A loadable particle comprising an energy donor as a first component and an energy acceptor as a second component positioned in said particle at an energy exchanging distance from one another, wherein the two components have a stokes shift of greater than or equal to 50 nm, wherein said first component has an excitation wavelength greater than approximately 550 nm and said second component has an emission wavelength greater than approximately 680 nm, said particle having bound on its surface, a protein, polypeptide, nucleic acid, nucleotide or protein containing ligand analogue.
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Abstract
Particles and methods for the detection or visualization of analytes using fluorescence energy transfer. Particles comprising an energy donor as a first component and a fluorescent dye as a second component positioned in said particles at an energy exchanging distance from one another, wherein the two components have a Stokes shift of greater than or equal to 50 nm, said particle having bound on its surface, a protein, polypeptide, nucleic acid, nucleotide or protein containing ligand analogue are disclosed and claimed.
63 Citations
57 Claims
- 1. A loadable particle comprising an energy donor as a first component and an energy acceptor as a second component positioned in said particle at an energy exchanging distance from one another, wherein the two components have a stokes shift of greater than or equal to 50 nm, wherein said first component has an excitation wavelength greater than approximately 550 nm and said second component has an emission wavelength greater than approximately 680 nm, said particle having bound on its surface, a protein, polypeptide, nucleic acid, nucleotide or protein containing ligand analogue.
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2. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned in said particle at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is phthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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3. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned in said particle at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said second component is phthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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4. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned in said particle at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is naphthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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5. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned in said particle at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said second component is naphthalocyanine and the two components have a stokes shift of greater than or equal to 50 nm.
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6. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is phthalocyanine substituted with at least one axial ligand and said second component is naphthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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7. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of trans-4-[4-(Dibutylamino)Styryl]-1-methyl pyridine and said second component is Silicon phthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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8. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of trans-4-[4-(Dibutylamino)Styryl]-1-methyl pyridine and said second component is Silion 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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9. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a salt of trans-4-[4-(Dibutylamino)Styryl]-1-methyl pyridine and said second component is a salt of 1,1-Dihexyl 3,3,3′
- ,3′
,-tetramethylindodicarbocyanine and the two components have a stokes shift of greater than or equal to 50 nm.
- ,3′
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10. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is Meso-tetra-2-aminophenyl porphine and said second component is Silicon phthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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11. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is Meso-tetra-2-aminophenyl porphine and said second component is a salt of 1,1-Dihexyl 3,3,3′
- ,3′
,-tetramethylindodicarbocyanine and the two components have a stokes shift of greater than or equal to 50 nm.
- ,3′
- 12. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is Meso-tetra-2-dimethylaminophenyl porphine and said second component is Silicon phthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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13. A particle comprising an energy donor as a first component and fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component a salt of 3-Ethyl-3′
- -ethyl carboxyethyl thiacarbocyanine and said second component is Silicon 2,3-Napthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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14. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 1,1′
- -Dioctadecyl-3,3,3′
,3′
-tetramethlyindodicarbocyanine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
- -Dioctadecyl-3,3,3′
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15. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 1,1′
- -Diethyl-3,3,3′
,3′
-tetramethylindodicarbocyanine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
- -Diethyl-3,3,3′
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16. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 1,1′
- -Dihexyl-3,3,3′
,3′
-tetramethlyindodicarbocyanine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
- -Dihexyl-3,3,3′
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17. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 3,3-Diethyl thiatricarbocyanine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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18. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 3,3-Dipropyl thiatricarbocyanine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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19. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 1,9-Dimethylmethylene blue and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide).
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20. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of N,N-Di(3-trimethylammoniumpropyl)thia-dicarbocyanine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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21. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 1,1′
- ,3,3,3′
,3′
-Hexamethylindotricarbocyanine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
- ,3,3,3′
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22. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of N-(3-Triethlylammoniumpropyl)-4-(4-(p-dibutylaminophenyl)butadienyl)pyridine and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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23. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is a salt of 1,1′
- ,3,3,3′
,3′
-Hexamethyl-4,4′
-5,5′
-dibenzo-2,2′
indotricarbocyanine and said second component is Silicon Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
- ,3,3,3′
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24. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is Fluorescein and said second component is Silicon Phthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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25. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is Chlorophyll and said second component is Silicon 2,3-Naphthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
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26. A particle loadable comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said first component is Chlorophyll and said second component is Silicon phthalocyanine bis(dimethylvinylsilyloxide) and the two components have a Stokes shift of greater than or equal to 50 nm.
- 27. A particle comprising an energy donor as a first component and second and third components comprising 2 fluorescent dyes positioned at an energy exchanging distance from one another, wherein said first component is a salt of trans-4-[4(Dibutylamino)styryl]-1-methyl pyridine and one of dyes is selected from the group consisting of Silicon phthalocyanine bis(dimethylpentafluorophenylsilyloxide) and Silicon phthalocyanine bis(dimethlylvinylsilyloxide).
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28. A particle comprising an energy donor as a first component and 3 fluorescent dyes positioned at an energy exchanging distance from one another, wherein said first component is a salt of trans-4-[4(Dibutylamino)styryl]-1-methyl pyridine and said three dyes are selected from the group consisting of Silicon phthalocyanine bis(trihexylsilyloxide), Silicon phthalacyanine bis(dimethylpentafluorophenylsilyloxide), Silicon phthalocyanine bis(dimethylvinylsilyloxide).
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29. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a styryl dye and said second component is phthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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30. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a styryl dye and said second component is naphthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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31. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a phenylbutadienyl dye and said second component is phthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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32. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a phenylbutadienyl dye and said second component is naphthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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33. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a phenylhexatrienyl dye and said second component is phthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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34. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a phenylhexatrienyl dye and said second component is naphthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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35. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a porphine dye and said second component is phthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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36. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a porphine dye and said second component is naphthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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37. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a carbocyanine dye and said second component is phthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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38. A loadable particle comprising an energy donor as a first component and a fluorescent dye as a second component positioned at an energy exchanging distance from one another, wherein said second component has an emission wavelength greater than approximately 680 nm, and wherein said first component is a carbocyanine dye and said second component is naphthalocyanine substituted with at least one axial ligand and the two components have a stokes shift of greater than or equal to 50 nm.
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48. A latex particle comprising an energy donor as a first component and an energy acceptor as a second component positioned in said particle at an energy exchanging distance from one another, wherein;
- (1) said first component has an excitation wavelength greater than approximately 550 nm and said second component has an emission wavelength greater than approximately 680 nm;
(2) said first component is a fluorescent dye and said second component is phthalocyanine-substituted with at least one axial ligand; and
(3) the two components have a Stokes shift of greater than or equal to 50 nm.
- (1) said first component has an excitation wavelength greater than approximately 550 nm and said second component has an emission wavelength greater than approximately 680 nm;
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49. A particle comprising an energy donor as a first component and an energy acceptor as a second component positioned in said particle at an energy exchanging distance from one another, wherein:
- (1) said first component has an excitation wavelength greater than approximately 550 nm and said second component has an emission wavelength, greater than approximately 680 nm;
(2) said second component is phthalocyanine substituted with at least one axial ligand; and
(3) the two components have a Stokes shift of greater than or equal to 50 nm.
- (1) said first component has an excitation wavelength greater than approximately 550 nm and said second component has an emission wavelength, greater than approximately 680 nm;
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50. A latex particle comprising an energy donor as a first component and an energy acceptor as a second component positioned in said particle at an energy exchanging distance from one another, wherein:
- (1) said first component is a fluorescent dye having an excitation wavelength greater than approximately 550 nm and said second component is a fluorescent dye having an emission wavelength greater than approximately 680 nm; and
(2) the two components have a Stokes shift of greater than or equal to 50 nm.
- (1) said first component is a fluorescent dye having an excitation wavelength greater than approximately 550 nm and said second component is a fluorescent dye having an emission wavelength greater than approximately 680 nm; and
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54. A particle for assaying analytes in a biological medium comprising an energy donor as a first component and an energy acceptor as a second component positioned in said particle at an energy exchanging distance from one another, wherein said first component has an excitation wavelength greater than about 600 nm, said second component has at least one emission wavelength other than the intrinsic fluorescent wavelength of the biological medium, and the two components have a Stokes shift of greater than or equal to 50 nm, said particle having bound on its surface, a protein, polypeptide, nucleic acid, ligand analogue, nucleotide or a protein-containing ligand analogue.
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55. A particle comprising an energy donor as a first component, an energy acceptor as a second component, and at least one additional energy acceptor as a third component, wherein:
- (1) said first component and said second component are positioned in said particle at an energy exchanging distance from one another;
(2) said first component has an excitation wavelength greater than approximately 550 nm and said second component has an emission wavelength greater than approximately 680 nm;
(3) said first component and said second component have a Stokes shift of greater than or equal to 50 nm; and
(4) said third component is a fluorescent dye, said third component exhibiting in the particle approximately the same excitation and emission wavelengths as said second component, whereby quenching is decreased and fluorescence intensity is increased by the combination of said second and said third component.
- (1) said first component and said second component are positioned in said particle at an energy exchanging distance from one another;
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56. A particle comprising an energy donor as a first component, an energy acceptor having an absorbance wavelength approximately equal to the emission wavelength of said first component as a second component, and at least one additional energy acceptor as a third component, wherein said first component and said second component are positioned in said particle at an energy exchanging distance from one another, wherein said first component and said second component have a Stokes shift of greater than or equal to 50 nm, and wherein said third component is a fluorescent dye exhibiting in the particle approximately the same excitation and emission wavelengths as said second component, whereby quenching is decreased and fluorescence intensity is increased by the combination of said second and said third component.
Specification