High-density fluorescent dye clusters
The present invention relates to modular sterically enhanced emission dye (SEED) clusters, wherein multiple SEED molecules are appended to a single polymeric chain.
1. A cluster comprising:
a 3 to 100 monomer unit polymer backbone, comprising two or more optionally substituted monomer units;
a solubilizing agent;
a conjugation site; and
a multiplicity of sterically enhanced emission dye molecules disposed along the polymer backbone.
2. The cluster of claim 1 , wherein at least two sterically enhanced emission dyes are attached to one or more of the monomer units.
3. The cluster of claim 1 , wherein the polymer is comprised of 10 to 40 monomer units.
4. The cluster of claim 1 wherein the hydrodynamic diameter of the cluster is less than 100 nm.
5. The cluster of claim 4 , wherein the hydrodynamic diameter of the cluster is between 2 and 15 nm.
6. The cluster of claim 1 , wherein each monomer unit is formed from a group independently selected from an alkenyl, an acrylate, an ether, an amine, or a combination thereof.
7. The cluster of claim 6 , wherein the monomer unit is formed from the group selected from acrylate, methylacrylate, vinyl, urethane, epoxide, or a combination thereof.
8. The cluster of claim 1 , wherein the solubilizing agent is selected from, oligo(ethylene glycol), oligosaccharides, cationically charged groups, anionically charged groups, zwitterionically charged groups, or combinations thereof.
9. The cluster of claim 1 , wherein each sterically enhanced emission dye is independently attached to the one or more monomer units either directly or through a linker.
10. The cluster of claim 1 , wherein each sterically enhanced emission dye within the cluster is the same.
11. The cluster of claim 1 , wherein at least two or more different sterically enhanced emission dyes are within the cluster.
12. The cluster of claim 11 , wherein the different sterically enhanced emission dyes have different absorption and/or emission wavelengths.
13. The cluster of claim 12 , wherein the wavelength is between 300 and 850 nm.
14. The cluster of claim 13 , wherein the molar ratio of different sterically enhanced emission dyes in each cluster is controlled.
15. The cluster of claim 1 , wherein each sterically enhanced emission dye is independently selected from the group consisting of rhodamines, fluoresceins, eosins, cyanines, boron dipyridyls, xanthenes, carbopyrinins, acridiniums, benzopyriniums, and acridiniums.
16. The cluster of claim 1 , wherein the molar ratio of the sterically enhanced emission dye and the solubilizing agents in each cluster is controlled.
17. The cluster of claim 1 , wherein the conjugation site is a reactive group selected from biotin, carboxyl, succinimidyl ester, sulpho-succinimidyl ester, N-hydroxysuccinimidyl ester, cadaverine, isothiocyanate, isocyanate, maleimide, haloacetamide acid halide, hydrazide, hydrazine, vinylsulphone, dichlorotriazine, phosphoramidite, sulphonyl halide, alkylimido ester, arylimido ester, carbodiimide, anhydrides or acyl azide; or a functional group selected from a primary amine, secondary amine, hydrazine derivatives, hydroxylamine derivatives, pyrazolone, sulphydryl, carboxyl, hydroxyl, thiol, imidazole, thiophosphate, or carbonyl including aldehyde or ketone.
18. The cluster of claim 17 , wherein the conjugation site is at the terminal end of the polymer or along the backbone of the polymer.
19. The cluster of claim 17 , wherein the conjugation site can bind to a target.
20. The cluster of claim 19 , wherein the target is a biological molecule.
21. A sustained emission high density luminescent cluster comprising:
a 3 to 100 monomer unit polymer backbone comprising two or more optionally substituted monomer units;
a solubilizing agent;
a conjugation site; and
a multiplicity of sterically enhanced emission dye molecules disposed along the polymer backbone such that rotational decay is suppressed and emission is sustained.