Radiative decay engineering
Compositions and methods for increasing the fluorescence intensity of molecules are provided. In particular, compositions and methods directed to increasing the intrinsic fluorescence of biomolecules and low quantum yield fluorophores are described. The intrinsic fluorescence of biomolecules is increased by positioning a metal particle and a biomolecule at a distance apart sufficient to increase the radiative decay rate of the biomolecule. Methods for the identification of nucleic acids are also provided. The compositions and methods can also be used to increase the emission of any fluorophore, such as the extrinsic probes used to label biomolecules.
1. A method for detecting a biomolecule by increasing intrinsic fluorescence of said biomolecule, said method comprising:
positioning a metal particle and said biomolecule at a distance sufficient to increase an intrinsic fluorescent emission from said biomolecule,
wherein said metal particle is a silver particle or a gold particle, and
wherein said biomolecule is selected from the group consisting of a nucleic acid; NADH; FAD; tryptophan, tyrosine, phenylalanine, proteins containing tryptophan, tyrosine, phenylalanine or any combination thereof; flavins; purines; pyrimidines; formycin; and phycobiliprotein;
exposing said biomolecule to an amount of exciting radiation sufficient to produce said intrinsic fluorescent emission for detection; and
detecting the intrinsic fluorescent emission from said biomolecule by a fluorescence detector.
2. A method for manipulating intrinsic fluorescent intensity of a biomolecule, said method comprising:
increasing a rate of radiative decay of the biomolecule by positioning a metal particle and the biomolecule at a distance sufficient to increase said radiative decay, said increase in said rate of radiative decay being associated with an increase in said intrinsic fluorescent intensity by intrinsic fluorescent emission of said biomolecule,
wherein said metal particle is a silver particle or a gold particle, and
wherein said biomolecule is selected from the group consisting of a nucleic acid; NADH; FAD; tryptophan, tyrosine, phenylalanine, proteins containing tryptophan, tyrosine, phenylalanine or any combination thereof; flavins; purines; pyrimidines; formycin; and phycobiliprotein;
exposing the biomolecule to exciting radiation; and
detecting an increased rate of radiative decay from said biomolecule by frequency-domain analysis of a fluorescent spectrum detected by a fluorescence detector.
3. The method of claim 1 , wherein said distance between said metal particle and said biomolecule is obtained by any of a chemical linker comprising an atom or a molecule, electromagnetic forces, charge fields, and gravity.
4. The method of claim 1 , wherein said biomolecule, when positioned at a distance greater than 2000 Å from said metal particle, has a quantum yield of less than 0.8.
5. The method of claim 1 , wherein said distance separating said biomolecule and said metal particle is about 50 Å to about 2000 Å.
6. The method of claim 1 , wherein said metal particle is suspended in a colloid.
7. The method of claim 1 , wherein said silver particle spherical dimensions of about 40 to 50 nm.
8. The method of claim 2 , wherein said distance between said metal particle and said biomolecule is obtained by any of a chemical linker comprising an atom or a molecule, electromagnetic forces, charge fields, and gravity.
9. The method of claim 2 , wherein said biomolecule, when positioned at a distance greater than 2000 Å from said metal particle, has a quantum yield of less than 0.8.
10. The method of claim 2 , wherein said distance separating said biomolecule and said metal particles is about 50 Å to about 2000 Å.
11. The method of claim 2 , wherein said metal particle is suspended in a colloid.
12. The method of claim 2 , wherein said silver particle has spherical dimensions of about 40 to 50 nm.