Fluorescent microparticles with controllable enhanced stokes shift

Claim 2

A fluorescent microparticle made by the process comprising: a) selecting a series of dyes comprising an initial donor dye with a desired excitation peak and a final acceptor dye with a desired emission peak, wherein the desired excitation peak and the desired emission peak are determined in a polymeric material, and wherein said polymeric material is made up of polymerizable monomers, and wherein each dye in the series has a spectral overlap sufficient to allow for significant energy transfer of excitation energy to the final acceptor dye, b) incorporating said series of dyes in a polymeric microparticle comprising said polymeric material, wherein said series of dyes are randomly incorporated into said polymeric microparticle.

Claim 3

A microparticle, as claimed in claim 1, wherein the spectral overlap allows for energy transfer that is greater than about 90%.

Claim 4

A microparticle, as claimed in claim 1, wherein the polymeric microparticle is polystyrene, brominated polystyrene, polyacrylic acid, polyacrylonitrile, polyacrylamide, polyacrolein, polydimethylsiloxane, polybutadiene, polyisoprene, polyurethane, polyvinylacetate, polyvinylchloride, polyvinylpyridine, polyvinylbenzylchloride, polyvinyltoluene, polyvinylidene chloride, or polydivinylbenzene.

Claim 5

A microparticle, as claimed in claim 1, wherein the fluorescent dyes are polyazaindacene; coumarin; oxazole or oxadiazole; aryl- or heteroaryl-substituted polyolefin wherein the olefin portion has 2-8 carbons; carbocyanine; phthalocyanine; oxazine; carbostyryl; or porphyrin dyes; or combinations thereof.

Claim 6

A microparticle, as claimed in claim 4, wherein at least one of the fluorescent dyes is a polyazaindacene dye of the formula: ##STR2## wherein R 1 -R 6 , which may be the same or different, are hydrogen, halogen, or alkyl, alkoxy, alkenyl, cycloalkyl, arylalkyl, acyl, wherein the alkyl portions each contain fewer than about 20 carbons, or aryl, heteroaryl, alone or in combination; and R 7 is nitrogen, methine, or halogen-, alkyl-, alkyl-, alkoxy-, alkenyl-, cycloalkyl-, arylalkyl-, acyl-, (the alkyl portions of which each contain fewer than about 20 carbons) aryl-, or heteroaryl-substituted methine.

Claim 7

A microparticle, as claimed in claim 1, wherein at least one of the fluorescent dyes is a polyazaindacene dye of the formula: ##STR3## wherein R 1 -R 6 , which may be the same or different, are hydrogen, halogen, or alkyl, alkoxy, alkenyl, cycloalkyl, arylalkyl, acyl, wherein the alkyl portions each contain fewer than about 20 carbons, or aryl, heteroaryl, alone or in combination; and R 7 is nitrogen, methine, or halogen-, alkyl-, alkoxy-, alkenyl-, cycloalkyl-, arylalkyl-, acyl-, (the alkyl portions of which each contain fewer than about 20 carbons) aryl-or heteroaryl-substituted methine.

Claim 8

A microparticle, as claimed in claim 6, wherein the spectral overlap allows for energy transfer that is greater than about 95%.

Claim 9

A microparticle, as claimed in claim 1, wherein the series of fluorescent dyes contains less than about 6 dyes in a total dye concentration that is between about 0.5% and 2% weight to weight.

Claim 10

A microparticle, as claimed in claim 1, wherein the ratio of initial donor to ultimate acceptor is between about 1:5 and about 10:1.

Claim 11

A microparticle, as claimed in claim 1, wherein the polymeric microparticle is polystyrene, polymethylmethacrylate, polyacrylonitrile, polyacrylamide, or polyacrolein; the series of fluorescent dyes contains less than about 6 dyes which are different polyazaindacene dyes of the formula: ##STR4## wherein R 1 -R 6 , which may be the same or different, are hydrogen, halogen, or alkyl, alkoxy, alkenyl, cycloalkyl, arylalkyl, acyl, (the alkyl portions of which each contain fewer than about 20 carbons) or aryl, heteroaryl, alone or in combination; and R 7 is nitrogen, methine, or halogen-, alkyl-, alkoxy-, alkenyl-, cycloalkyl-, arylalkyl-, acyl-, (the alkyl portions of which each contain fewer than about 20 carbons) aryl- or heteroaryl-substituted methine; for which the total dye concentration is between about 0.8 and 1.2% (w/w); and the ratio between initial donor and ultimate acceptor dyes is between about 4:1 and about 6:1.

Claim 12

A fluorescent microparticle comprising: a series of fluorescent dyes having an initial donor dye with a desired excitation peak and an ultimate acceptor dye with a desired emission peak, wherein the desired excitation peak and the desired emission peak are determined in a polymeric material, wherein said polymeric material is made up of polymerizable monomers, and wherein each dye in the series has a spectral overlap sufficient to allow for significant energy transfer of excitation energy to the ultimate acceptor dye, wherein said series of dyes is randomly incorporated in a polymeric microparticle comprising said polymeric material such that the total dye concentration is less than about 10% w/w and the ratio of initial donor to ultimate acceptor is between about 1:5 and about 1:10.

Claim 13

A fluorescent microparticle comprising: a series of fluorescent dyes having an initial donor dye with a desired excitation peak and an ultimate acceptor dye with a desired emission peak, wherein the desired excitation peak and the desired emission peak are determined in a polymeric material, wherein said polymeric material is made up of polymerizable monomers, and wherein each dye in the series has a spectral overlap sufficient to allow for significant energy transfer of excitation energy to the ultimate acceptor dye, wherein said series of dyes is randomly incorporated in a polymeric microparticle comprising said polymeric material such that the total dye concentration is less than about 10% w/w and the ratio of initial donor to ultimate acceptor is between about 1:5 and about 1:10.

Claim 14

A microparticle, as claimed in claim 11, wherein the microparticle is polystyrene, brominated polystyrene, polyacrylic acid, polyacrylonitrile, polyacrylamide, polyacrolein, polydimethylsiloxane, polybutadiene, polyisoprene, polyurethane, polyvinylacetate, polyvinylchloride, polyvinylpyridine, polyvinylbenzylchloride, polyvinyltoluene, polyvinylidene chloride, or polydivinylbenzene and is less than about 15 micrometers in diameter.

Claim 15

A microparticle, as claimed in claim 12, wherein the series of dyes contains less than about 6 polyazaindacene dyes of the formula: ##STR5## wherein R 1 -R 6 , which may be the same or different, are hydrogen, halogen, or alkyl, aldoxy, alkenyl, cycloalkyl, arylaklyl, acryl, wherein the alkyl portions each contain fewer than about 20 carbons, or aryl, heteroaryl, alone or in combination; and R 7 is nitrogen, methine, or halogen-, alkyl-, alkoxy-, alkenyl-, cycloalkyl-, arylalkyl-, acyl-, (the alkyl portions of which each contain fewer than about 20 carbons) aryl-or heteroarylsubstituted methine.

Claim 16

A microparticle, as claimed in claim 13, further comprising a bioreactive substance, wherein said bioreactive substance has been either bound covalently or passively adsorbed.

Claim 17

A microparticle, as claimed in claim 14, wherein said bioreactive substance is a biomolecule that is biotin, avidin, streptavidin, digoxigenin, or a nucleic acid.