Fluorescent polymeric materials containing lipid soluble rhodamine dyes

• US 20040225037A1
• Filed: 05/04/2004
• Published: 11/11/2004
• Est. Priority Date: 05/09/2003
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Menchen, Steven M., Benson, Scott C., Zou, Ruiming, Lam, Joe Y.L.
• Granted Patent: US 7,432,298 B2
• Time in Patent Office: Days
• US Class Current: 524/90
• CPC Class Codes: C07D 311/80 Dibenzopyrans; Hydrogenated dibenzopyrans
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C09B 67/0092 Dyes in solid form
• CPC Class Codes: C09B 67/0097 Dye preparations of special physical nature; Tablets, films, extrusion, microcapsules, sheets, pads, bags with dyes
• CPC Class Codes: C09B 69/103 containing a diaryl- or triarylmethane dye
• CPC Class Codes: C09K 11/025 non-luminescent particle coatings or suspension media
• CPC Class Codes: C09K 11/06 containing organic luminescent materials
• CPC Class Codes: C09K 2211/1088 containing oxygen as the only heteroatom
• CPC Class Codes: C09K 2211/1416 Condensed systems
• CPC Class Codes: C09K 2211/145 containing oxygen as the only heteroatom

Abstract

Fluorescent polymeric materials are disclosed comprising a polymer and one or more lipid soluble rhodamine dyes. The materials are especially useful in the preparation of multicolored microparticles, especially multicolored polystyrene microparticle, for use in the multiplexed analysis of a plurality of analytes in a single sample. When excited by a light source, the materials give off a unique emission based on the nature, concentration and ratio of the dyes therein. Methods of preparing and using said materials are also disclosed.

Claims

1. A fluorescent polymeric material comprising a polymer and at least one lipid-soluble rhodamine dye comprising one of the following core structures:

5. The fluorescent polymeric material of any one of claims 1-4 wherein at least one of R^3′

, R^3″

, R^6″

and R^6′

is a lipophilic substituent selected from (C4-C20) alkyls and heteroalkyls, (C5-C20) aryls and heteroaryls, and (C6-C40) arylalkyls or heteroaryl alkyls.

6. The fluorescent polymeric material of any one of claims 1-4 wherein at least one of R^3′

, R^3″

, R^6″

and R^6′

is a lipophilic substituent is selected from (C7-C20) alkyls.

7. The fluorescent polymeric material of any one of claims 1-4 wherein at least two of R^3′

, R^6″

, R^3″

and R^6″

are the same or different lipophilic substituent.

8. The fluorescent polymeric material of any one of claims 1-4 wherein all of R^3′

, R^6″

, R^3″

and R^6″

are the same or different lipophilic substituent.

9. The fluorescent polymeric material of any one of claims 1-4 wherein the substituents in core structures (I), (II), (III) and (IV) are further defined as follows:

(i.) with respect to core structure (I),

R¹ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R¹ is taken together with R² form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R² is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R² is taken together with Rto form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R² is taken together with R^3′

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^3′

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^3′

is taken together with R² to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^3″

is selected from (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^3″

is taken together with R⁴ to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁴ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively R⁴ is taken together with R^3″

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁵ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable 10 group or, alternatively R⁵ is taken together with R⁶ to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^6″

is selected from (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups or, alternatively R⁶ is taken together with R⁵ to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^6′

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^6′

is taken together with R⁷ to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁷ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or R^b groups, or, alternatively, R⁷ is taken together with R^6′

to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁷ is taken together with R⁸ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁸ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁸ together with R⁷ form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R¹¹ and R¹⁵ are each, independently of one another, selected from hydrogen, halo, (C1-C20) alkyl, haloalkyl, —

OR^y, —

SR^y, —

SOR^y, —

SO₂R^y, —

C(O)OR^y, —

S(O)₂OR^y, amide, sulfonamide and nitrile,

R¹², R¹¹ and R¹⁴ are each, independently of one another, selected from hydrogen R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^x is selected from —

NR^cR^c, —

OR^d, —

SR^d, halo, haloalkyl, —

CN, —

NC, —

OCN, —

SCN, —

NO, —

NO₂, —

N₃, —

S(O)R^d, —

S(O)₂R^d, —

S(O)₂OR^d, —

S(O)NR^cR ^c, —

S(O)₂NR^cR^c, —

SO(O)R^d, —

SO(O)₂R^d, —

SO(O)₂OR^d, —

SO(O)₂NR^cR^c, —

C(O)R^d, —

C(O)O R^d, —

C(O)NR^cR^c, —

C(NH)NR^cR^c, —

OC(O)R^d, —

OC(O)OR^d, —

OC(O)NR^cR^c and —

OC(NH)NR^cR^c,

R^y is selected from (C1-C20) alkyls or heteroalkyls optionally substituted with lipophilic substituents, (C5-C20) aryls or heteroaryls optionally substituted with lipophilic substituents and (C6-C26) arylalkyl or heteroarylalkyls optionally substituted with lipophilic substituents,

R^a is selected from hydrogen, (C1-C8) alkyl or heteroalkyl, (C5-C20) aryl or heteroaryl and (C6-C28) arylalkyl or heteroarylalkyl,

R^b is selected from —

NR^cR^c, ═

O, —

OR^d, ═

S, —

SR^d, ═

NR^d, ═

NOR^d, halo, haloalkyl, —

CN, —

NC, —

OCN, —

SCN, —

NO, —

NO₂, ═

N₂, —

N₃, —

S(O)R^d, —

S(O)₂R ^d, —

S(O)₂OR^d, —

S(O)NR^cR^c, —

S(O)₂NR^cR^c, —

SO(O)R^d, —

SO(O)₂R^d, —

SO(O)₂OR^d, —

SO(O)₂NR^cR^c, —

C(O)R^d, —

C(O)OR^d, —

C(O)NR^cR^c, —

C(NH)NR^cR^c, —

OC(O)R^d, —

OC(O)OR^d, —

OC(O)NR^cR^c and —

OC(NH)NR^cR^c,

each R^c is independently hydrogen or R^d, or, alternatively, each R^c is taken together with the nitrogen atom to which it is bonded to form a 5 to 8-membered saturated or unsaturated ring which may optionally include one or more of the same or different additional heteroatoms and which may optionally be substituted with one or more of the same or different R^a or R^d groups,

each R^d is independently R^a or R^a substituted with one or more of the same or different R^a or R^e groups, and

each R^e is selected from —

NR^aR^a, ═

O, —

OR^a═

S, —

SR^a, ═

NR^a, ═

NOR^a, halo, haloalkyl, —

CN, —

NC, —

OCN, —

SCN, —

NO, —

NO₂, ═

N₂, —

N₃, —

S(O)R^a, —

S(O)₂R^a, —

S(O)₂OR^a, —

S(O)NR^aR^a, —

S(O)₂NR^aR^a, —

SO(O)R^a, —

SO(O)₂R^a, —

SO(O)₂OR^a, —

SO(O)₂NR^aR^a, —

C(O)R^a, —

C(O)OR^a, —

C(O)NR^aR^a, —

C(NH)NR^aR^a, —

OC(O)R^a, —

OC(O)OR^a, —

OC(O)NR^aR^a and —

OC(NH)NR^aR^a; and

(ii.) with respect to core structure (II),

R¹, R², R^3′

, R^3″

, R⁴, R⁵, R^6″

, R^6′

, R⁷, R⁸, R^x, R^a, R^b, R^c, R^d, and R^e are as first defined in section 5.6 with respect to core structure (I), and

R⁹ is selected from hydrogen alkyl, halo, haloalkyl, or nitrile;

(iii.) with respect to core structure (III),

R¹, R^3″

, R⁵, R^6″

, R^6′

, R⁷, R⁸, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R^x, R^y, R^a, R^b, R^c, R^d, and R^e are as first defined in section 5.6 with respect to core structure (I),

R² is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R² is taken together with Ror R^2′

to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^2′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^2′

is taken together with R² to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^2′

is taken together with R^3′

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^3′

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^3′

is taken together with R^2′

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁴ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively R⁴ is taken together with R^3″

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁴ is taken together with R^4′

to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, and

R^4′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively R^4′

is taken together R⁴ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups; and

(iv.) with respect to core structure (IV),

R¹, R^3″

, R^6″

, R⁸, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R^x, R^y, R^a, R^b, R^c, R^d, and R^e areas first defined in section 5.6 with respect to core structure (I),

R², R^2′

, R^3′

, R⁴ and R^4′

are as defined in section 5.8 with respect to core structure (II),

R^5′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b group or, alternatively R⁵ is taken together with R⁵ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁵ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b group or, alternatively R⁵ is taken together with R^6″

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁵ is taken together with R^5′

to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^6″

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^6′

is taken together with R^7′

to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^7′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or R^b groups, or, alternatively, R^7″

is taken together with R^6′

to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^7′

is taken together with R⁷ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, and

R⁷ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or R^b groups, or, alternatively, R⁷ is taken together with R^7′

or R⁸ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups.

10. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (I).

11. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (I) and, additionally, the substituents therein comprise one or more of the following features:

R¹ and R⁸ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, halo, and —

CF₃, or are part of a fused aryl or heteroaryl group with R² and R⁷, respectively;

R² and R⁷ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, halo, —

CF₃, —

CN, carboxylate, ester, sulfone, amino, amido and oxoether, or are part of a fused aryl or heteroaryl group with R¹ and R⁸, respectively, or are part of a 5 or 6 membered ring with R^3′

and R^6′

, respectively;

R^3′

and R^6′

are each, independently of one another, selected from hydrogen and (C4-C20) alkyl, or are part of a 5 or 6 membered ring with R² and R⁷, respectively;

R^3″

and R^6″

are each, independently of one another, selected from hydrogen and (C4-C20) alkyl or are part of a 5 or 6 membered ring with R⁴ and R⁵, respectively;

at least two of R^3′

, R^3″

, R^6′

, R^6″

are not hydrogen;

R⁴ and R⁵ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, halo, —

CF₃, aryl, heteroaryl, —

CN, carboxylate, ester, and sulfone, or are part of a 5 or 6 membered ring with R^3″

and R^6″

, respectively;

R¹¹ and R¹⁵ are each, independently, selected from hydrogen, halo, ester, amide, sulfonamide, (C1-C20) alkyl, —

OR^e, —

SR^e, —

CN and sulfone; and

R¹², R¹³ and R¹⁴ are each, independently of one another, selected from hydrogen, halo, alkyl, aryl, (C4 to C20) alkyl ester, sulfone, —

CF₃ and —

CN.

12. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (I) and, additionally, the substituents therein comprise one or more of the following features:

R¹ and R⁸ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, and halo, or are part of a fused aryl or heteroaryl group with R² and R⁸, respectively;

R² and R⁷ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, and halo, or are part of a fused aryl or heteroaryl group with R¹ and R⁸ respectively, or are part of a 5 or 6 membered ring with R^3′

and R^6′

, respectively;

R^3′

and R^6′

are each the same or different (C4-C20) alkyl;

R^3″

and R^6″

are each, independently of one another, selected from hydrogen and the same or different (C4-C20) alkyl, or are part of a 5 or 6 membered ring with R⁴ and R⁵, respectively;

R⁴ and R⁵ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, aryl and heteroaryl and halo, or are part of a 5 or 6 membered ring with R^3″

and R^6″

, respectively;

R¹¹ and R¹⁵, independently of one another, selected from hydrogen, fluoro, chloro, (C1-C20) alkyl, —

OR^e and —

SR^e; and

R¹², R¹³ and R¹⁴ are each, independently of one another, selected from hydrogen and halo.

13. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises one of the following structural embodiments of core structure (I):

14. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye is one of the following embodiments of core structure (I):

15. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (II).

16. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (II) and, additionally, the substituents therein comprise one or more of the following features:

R¹ when taken together with R² forms part of a fused benzo, naptho or polycyclic aryleno group that is optionally substituted with one or more of the same or different R^a or suitable R^b groups;

R⁷ when taken together with R⁸ forms part of a fused benzo, naptho or polycyclic aryleno group that is optionally substituted with one or more of the same or different R^a or suitable R^b groups; and

R⁹ is hydrogen.

17. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises one of the following structural embodiments of core structure (II):

18. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye is selected from the following embodiments of core structure (II):

19. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (III).

20. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (III) and further comprises one or more substituted or unsubstituted five or six membered fused rings that comprise, as part of the ring, a nitrogen atom from either the exocyclic amine or exocyclic imminium group and one of R^2′

, R⁴, R⁵ or R⁷.

21. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (IV).

22. The fluorescent polymeric material of claim 9 wherein the lipid soluble rhodamine dye comprises core structure (IV) and further comprises one or more substituted or unsubstituted five or six membered fused rings that comprise, as part of the ring, a nitrogen atom from either the exocyclic amine or exocyclic imminium group and one of R^2′

, R⁴, R⁵ or R^7′

.

23. The fluorescent polymeric material of any one of claims 1-3 in which the polymer is formed by polymerizing one or more of the following monomers:

substituted or unsubstituted styrenes, acrylate, alkyl acrylates, methacrylate, alkyl methacrylates, acrylonitrile, alkyl acrylonitrile, esters, acetates, amides, alcohols, acrolein, dimethylsiloxane, butadiene, isoprene, urethane, vinylacetate, vinylchloride, vinylpyridine, vinylbenzylchloride, vinyltoluene, vinylidene chloride and divinylbenzene, and mixtures thereof, with or without the additional presence of a crosslinking agent.

24. The fluorescent polymeric material of claim 23 wherein the polymer is a hydrophobic copolymer of styrene, acrylic acid, and a polyvinyl crosslinking agent.

25. The fluorescent polymeric material of any one of claims 1-4 wherein the polymer is in the form of a microparticle having a diameter in the range of about 0.01 to 1000 micrometers.

26. The fluorescent polymeric material of any one of claims 1-4 in which the lipid-soluble rhodamine dye is incorporated into the polymer non-covalently.

27. The fluorescent polymeric material of any one of claims 1, 3 and 4 wherein the polymer is activated so that it comprises one or more groups that have a reactive functionality.

28. The fluorescent polymeric material of any one of claim 27 wherein the reactive functionality is an electrophile selected from pentafluorphenyl ester, NHS-ester, acrylamides, acyl azides, acyl halides, acyl nitriles, aldehydes or ketones, alkyl halides, alkyl sulfonates, anydrides, aryl halides, aziridines, boronates, carboxylic acids, carbodiimides, diazoalkenes, epoxides, haloacetamides, halotriazines, imido esters, isocyanates, isothiocyanates, maleimides, phorphoramidites, silyl halides, sulfonate esters, and sulfonyl halides.

29. The fluorescent polymeric material of claim 27 wherein the reactive functionality is a nucleophile selected from amines/anilines, alcohols/phenols, thiols, hydrazines, and hydroxylamines.

30. The fluorescent polymeric material of any one of claims 1-4 that further includes another substance that is absorbed into, adsorbed onto or covalently attached to the polymer.

31. The fluorescent polymeric material of claim 30 in which the other substance is a biomolecule or a probe for a biomolecule.

32. The fluorescent polymeric material of claim 31 in which the other substance is a biomolecule selected from amino acids, polypeptides, glycosolated and unglycosolated proteins, nucleosides, nucleotides, oligonucleotides, polynucleotides, nucleic acids, polynucleic acids, carbohydrates, and steroids.

33. The fluorescent polymeric material of claim 31 in which the other substance is a biomolecule selected from haptens, antibodies, enzymes, toxins and hormones.

34. The fluorescent polymeric material of claim 30 in which the other substance is selected from antigens, drugs, vitamins, poisons, and pathogens.

35. The fluorescent polymeric material of any one of claims 1-4 which includes a mixture of from 1 to 10 fluorescent dyes.

36. The fluorescent polymeric material of claim 35 in which each of the fluorescent dyes is independently selected lipid soluble rhodamine dye comprising one of core structures (I), (II), (III) and (IV), acridine dyes, alizarene dyes, azo dyes, anthraquinine dyes, bodipy dyes, coumarin dyes, cyaniine dyes, fluorescein dyes, lanthanide complexes, oxazine dyes, phenazathionium dyes, phenazoxonium dyes, porphyorin dyes, pyrene dyes, pyrilium dyes, perylene dyes, phenoxazine dyes, phenezine dyes, rhodol dyes and xanthene dyes.

37. The fluorescent polymeric material of claim 35 in which all of the fluorescent dyes fluoresce when irradiated with the same excitation source.

38. The fluorescent polymer material of claim 35 in which two or more fluorescent dyes fluoresce when irradiated with multiple excitation sources.

39. The fluorescent polymeric material of claim 35 wherein at least one of the fluorescent dyes is an energy transfer dye.

2. A fluorescent polymeric material comprising a polymer activated with functional groups suitable for covalent attachment of molecules thereto, said polymer having incorporated internally therein at least one lipid-soluble rhodamine dye comprising one of the following core structures:

3. A fluorescent polymeric material comprising a polymeric particle that swells in organic solvent, said polymeric particle having incorporated internally therein at least one lipid-soluble rhodamine dye comprising one of the following core structures:

4. A fluorescent polymeric material comprising a polymeric particle formed from a crosslinked or uncrosslinked polymer selected from polystyrene homopolymer and copolymers of styrene polymerized with one or more other monomers, said polymeric particle having incorporated internally therein at least one lipid-soluble rhodamine dye comprising one of the following core structures:

40. A collection of fluorescent polymeric particle populations, each of which has a unique, spectrally resolvable fluorescent emission spectrum and at least one of which comprises particles that comprise a polymer having incorporated internally therein at least one lipid-soluble rhodamine dye comprising one of the following core structures:

41. The collection of claim 40 wherein at least one of R^3′

, R^3″

, R^6″

and R^6′

is a lipophilic substituent selected from (C4-C20) alkyls and heteroalkyls, (C5-C20) aryls and heteroaryls, and (C6-C40) arylalkyls or heteroaryl alkyis.

42. The collection of claim 40 wherein at least one of R^3′

, R^3″

, R^6″

and R^6′

is a lipophilic substituent is selected from (C7-C20) alkyls.

43. The collection of claim 40 wherein at least two of R^3′

, R^6″

, R^3″

and R^6″

are the same or different lipophilic substituent.

44. The collection of claim 40 wherein all of R^3′

, R^6″

, R^3″

and R^6″

are the same or different lipophilic substituent.

45. The collection of claim 40 wherein the substituents in core structures (I), (II), (III) and (IV) are further defined as follows:

(i.) with respect to core structure (I),

R² is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R¹ is taken together with R² form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R² is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R² is taken together with R¹ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R² is taken together with R³ to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^3′

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^3′

is taken together with R² to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^3″

is selected from (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl, alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^3″

is taken together with R⁴ to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁴ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively R⁴ is taken together with R³ to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁵ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b group or, alternatively R⁵ is taken together with R^6″

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, R^6″

is selected from (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups or, alternatively R^6″

is taken together with R⁵ to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^6′

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^6′

is taken together with R⁷ to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁷ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or R^b groups, or, alternatively, R⁷ is taken together with R^6′

to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁷ is taken together with R⁸ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁸ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁸ together with R⁷ form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R¹¹ and R¹⁵ are each, independently of one another, selected from hydrogen, halo, (C1-C20) alkyl, haloalkyl, —

OR^y, —

SR^y, —

SOR^y, —

SO₂OR^y, —

C(O)OR^y, —

S(O)₂OR^y, amide, sulfonamide and nitrile,

R¹², R¹³ and R¹⁴ are each, independently of one another, selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^x is selected from —

NR^cR^c, —

OR^d, —

SR^d, halo, haloalkyl, —

CN, —

NC, —

OCN, —

SCN, —

NO, —

NO₂, —

N₃, —

S(O)R^d, —

S(O)₂R^d, —

S(O)₂OR^d, —

S(O)NR^cR ^c, —

S(O)₂NR^cR^c, —

SO(O)R^d, —

SO(O)₂R^d, —

SO(O)₂OR^d, —

SO(O)₂NR^cR^c, —

C(O)R^d, —

C(O)O R^d, —

C(O)NR^cR^c, —

C(NH)NR^cR^c, —

OC(O)R^d, —

OC(O)OR^d, —

OC(O)NR^cR^c and —

OC(NH)NR^cR^c,

R^y is selected from (C1-C20) alkyls or heteroalkyls optionally substituted with lipophilic substituents, (C5-C20) aryls or heteroaryls optionally substituted with lipophilic substituents and (C6-C26) arylalkyl or heteroarylalkyls optionally substituted with lipophilic substituents,

R^a is selected from hydrogen, (C1-C8) alkyl or heteroalkyl, (C5-C20) aryl or heteroaryl and (C6-C28) arylalkyl or heteroarylalkyl,

R^b is selected from —

NR^cR^c, ═

O, —

OR^d, ═

S, —

SR^d, ═

NR^d, ═

NOR^d, halo, haloalkyl, , —

CN, —

NC, —

OCN, —

SCN, —

NO, —

NO₂, ═

N₂, —

N₃, —

S(O)R^d, —

S(O)₂R ^d, —

S(O)₂OR^d, —

S(O)NR^cR^c, —

S(O)₂NR^cR^c, —

SO(O)R^d, —

SO(O)₂R^d, —

SO(O)₂OR^d, —

SO(O)₂NR^cR^c, —

C(O)R^d, —

C(O)OR^d, —

C(O)NR^cR^c, —

C(NH)NR^cR^c, —

OC(O)R^d, —

OC(O)OR^d, —

OC(O)NR^cR^c and —

OC(NH)NR^cR^c,

each R^c is independently hydrogen or R^d, or, alternatively, each R^c is taken together with the nitrogen atom to which it;

is bonded to form a 5 to 8-membered saturated or unsaturated ring which may optionally include one or more of the same or different additional heteroatoms and which may optionally be substituted with one or more of the same or different R^a or R^d groups,

each R^d is independently R^a or R^a substituted with one or more of the same or different R^a or R^e groups, and

each R^e is selected from —

NR^aR^a, ═

O, —

OR^a, ═

S, —

SR^a, ═

NR^a, ═

NOR^a, halo, haloalkyl, —

CN, —

NC, —

OCN, —

SCN, —

NO, —

NO₂, ═

N₂, —

N₃, —

S(O)R^a, —

S(O)₂R^a, —

S(O)₂OR^a, —

S(O)NR^aR^a, —

S(O)₂NR^aR^a, —

SO(O)R^a, —

SO(O)₂R^a, —

SO(O)₂OR^a, —

OS(O)₂NR^aR^a, —

C(O)R^a, —

C(O)OR^a, —

C(O)NR^aR^a, —

C(NH)NR^aR^a, —

OC(O)R^a, —

OC(O)OR^a, —

OC(O)NR^aR^a and —

OC(NH)NR^aR^a; and

(ii.) with respect to core structure (II),

R¹, R², R^3′

, R^3″

, R⁴, R⁵, R^6″

, R^6′

, R⁷, R⁸, R^x, R^a, R^b, R^c, R^d, and R^e are as first defined in section 5.6 with respect to core structure (I), and

R⁹ is selected from hydrogen, alkyl, halo, haloalkyl, or nitrile;

(iii.) with respect to core structure (III),

R¹, R^3″

, R⁵, R^6″

, R^6′

, R⁷, R⁸, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R^x, R^y, R^a, R^b, R^c, R^d, and R^e are as first defined in section 5.6 with respect to core structure (I),

R² is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R² is taken together with R¹ or R^2′

to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^2′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^2′

is taken together with R² to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^2′

is taken together with R^3′

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^3′

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^3′

is taken together with R^2′

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁴ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively R⁴ is taken together with R^3″

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁴ is taken together with R^4′

to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, and

R^4′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively R^4′

is taken together R⁴ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups; and

(iv.) with respect to core structure (IV),

R¹, R^3″

, R^6″

, R⁸, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R^x, R^y, R^a, R^b, R^c, R^d, and R^e are as first defined in section 5.6 with respect to core structure (I),

R², R^2′

, R^3′

, R⁴, and R^4′

are as defined in section 5.8 with respect to core structure (III),

R^5′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b group or, alternatively R⁵ is taken together with R⁵ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R⁵ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b group or, alternatively R⁵ is taken together with R^6″

to form a 5- or 6-membered ring which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R⁵ is taken together with R^5′

to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^6′

is selected from hydrogen, (C4-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroarylalkyl optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^6′

is taken together with R^7′

to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups,

R^7′

is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or R^b groups, or, alternatively, R^7′

is taken together with R^6′

to form a 5- or 6-membered ring optionally substituted with one or more of the same or different R^a or suitable R^b groups, or, alternatively, R^7′

is taken together with R⁷ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups, and

R⁷ is selected from hydrogen, R^x, (C1-C20) alkyl or heteroalkyl optionally substituted with one or more of the same or different R^b groups, (C5-C20) aryl or heteroaryl optionally substituted with one or more of the same or different R^a or suitable R^b groups and (C6-C40) arylalkyl or heteroaryl alkyl optionally substituted with one or more of the same or different R^a or R^b groups, or, alternatively, R⁷ is taken together with R^7′

or R⁸ to form part of a benzo, naptho or polycyclic aryleno group which is optionally substituted with one or more of the same or different R^a or suitable R^b groups.

46. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (I).

47. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (I) and, additionally, the substituents therein comprise one or more of the following features:

R¹ and R⁸ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, halo, and —

CF₃, or are part of a fuised aryl or heteroaryl group with R² and R⁷, respectively;

R² and R⁷ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, halo, —

CF₃, —

CN, carboxylate, ester, sulfone, amino, amido and oxoether, or are part of a fused aryl or heteroaryl group with R¹ and R⁸, respectively, or are part of a 5 or 6 membered ring with R^3′

and R^6′

, respectively;

R^3′

and R^6′

are each, independently of one another, selected from hydrogen and (C4-C20) alkyl, or are part of a 5 or 6 membered ring with R² and R⁷, respectively;

R^3″

and R^6″

are each, independently of one another, selected from hydrogen and (C4-C20) alkyl or are part of a 5 or 6 membered ring with R⁴ and R⁵, respectively;

at least two of R^3′

, R^3″

, R^6′

, R^6″

are not hydrogen;

R⁴ and R⁵ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, halo, —

CF₃, aryl, heteroaryl, —

CN, carboxylate, ester, and sulfone, or are part of a 5 or 6 membered ring with R^3″

and R^6″

, respectively;

R¹¹ and R¹⁵ are each, independently, selected from hydrogen, halo, ester, amide, sulfonamide, (C1-C20) alkyl, —

OR^e, —

SR^e, —

CN and sulfone; and

R¹², R¹³ and R¹⁴ are each, independently of one another, selected from hydrogen, halo, alkyl, aryl, (C4 to C20) alkyl ester, sulfone, —

CF₃ and —

CN.

48. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (I) and, additionally, the substituents therein comprise one or more of the following features:

R¹ and R⁸ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, and halo, or are part of a fuised aryl or heteroaryl group with R² and R⁸, respectively;

R² and R⁷ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, and halo, or are part of a fused aryl or heteroaryl group with R¹ and R⁸ respectively, or are part of a 5 or 6 membered ring with R^3′

and R^6′

, respectively;

R^3′

and R^6′

are each the same or different (C4-C20) alkyl;

R^3″

and R^6″

are each, independently of one another, selected from hydrogen and the same or different (C4-C20) alkyl, or are part of a 5 or 6 membered ring with R⁴ and R⁵, respectively;

R⁴ and R⁵ are each, independently of one another, selected from hydrogen, (C1-C8) alkyl, aryl and heteroaryl and halo, or are part of a 5 or 6 membered ring with R^3″

and R^6″

, respectively;

R¹¹ and R¹⁵, independently of one another, selected from hydrogen, fluoro, chloro, (C1-C20) alkyl, —

OR^e and —

SR^e; and

R¹², R¹³ and R¹⁴ are each, independently of one another, selected from hydrogen and halo.

49. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises one of the following structural embodiments of core structure (I):

50. The collection of claim 40 wherein the lipid soluble rhodamine dye is one of the following embodiments of core structure (I):

51. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (II).

52. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (II) and, additionally, the substituents therein comprise one or more of the following features:

R¹ when taken together with R² forms part of a fused benzo, naptho or polycyclic aryleno group that is optionally substituted with one or more of the same or different R^a or suitable R^b groups; and

R⁷ when taken together with R⁸ forms part of a fused benzo, naptho or polycyclic aryleno group that is optionally substituted with one or more of the same or different R^a or suitable R^b groups.

53. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises one of the following structural embodiments of core structure (II):

54. The collection of claim 40 wherein the lipid soluble rhodamine dye is one of the following embodiments of core structure (II):

55. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (III).

56. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (III) and further comprises one or more substituted or unsubstituted five or six membered fused rings that comprise, as part of the ring, a nitrogen atom from either the exocyclic amine or exocyclic imminium group and one of R^2′

, R⁴, R⁵ or R⁷.

57. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (IV).

58. The collection of claim 40 wherein the lipid soluble rhodamine dye comprises core structure (IV) and further comprises one or more substituted or unsubstituted five or six membered fused rings that comprise, as part of the ring, a nitrogen atom from either the exocyclic amine or exocyclic imminium group and one of R^2′

, R⁴, R⁵ or R^7′

.

59. The collection of claim 40 in which the polymer is formed from one or more of the following monomers:

substituted or unsubstituted styrenes, acrylate, alkyl acrylates, methacrylate, alkyl methacrylates, acrylonitrile, alkyl acrylonitrile, esters, acetates, amides, alcohols, acrolein, dimethylsiloxane, butadiene, isoprene, urethane, vinylacetate, vinylchloride, vinylpyridine, vinylbenzylchloride, vinyltoluene, vinylidene chloride and divinylbenzene, and mixtures thereof, with or without the additional presence of a crosslinking agent.

60. The collection of claim 59 in which the polymer is a hydrophobic copolymer of styrene, acrylic acid, and a polyvinyl crosslinking agent.

61. The collection of claim 40 in which each of the particles comprising the lipid soluble rhodamine dye is a microparticle having a diameter in the range of about 0.01 to 1000 micrometers.

62. The collection of claim 40 in which the lipid-soluble rhodamine dye is incorporated into the polymer non-covalently.

63. The collection of claim 40 in which one or more of the particles comprising the lipid soluble rhodamine dye is activated to comprise one or more groups that have a reactive functionality.

64. The collection of claim 63 in which the reactive functionality is an electrophile selected from pentafluorphenyl ester, NHS-ester, acrylamides, acyl azides, acyl halides, acyl nitriles, aldehydes or ketones, alkyl halides, alkyl sulfonates, anydrides, aryl halides, aziridines, boronates, carboxylic acids, carbodiimides, diazoalkenes, epoxides, haloacetamides, halotriazines, imido esters, isocyanates, isothiocyanates, maleimides, phorphoramidites, silyl halides, sulfonate esters, and sulfonyl halides.

65. The collection of claim 63 in which the reactive functionality is a nucleophile selected from amines/anilines, alcohols/phenols, thiols, hydrazines, and hydroxylamines.

66. The collection of claim 40 in which the particles comprising the lipid soluble rhodamine dye further include another substance that is absorbed into, adsorbed onto or covalently attached to the particles.

67. The collection of claim 66 in which the other substance is a biomolecule or a probe for a biomolecule.

68. The collection of claim 67 in which the other substance is a biomolecule selected from amino acids, polypeptides, glycosolated and unglycosolated proteins, nucleosides, nucleotides, oligonucleotides, polynucleotides, nucleic acids, polynucleic acids, carbohydrates, and steroids.

69. The collection of claim 67 in which the other substance is a biomolecule selected from haptens, antibodies, enzymes, toxins and hormones.

70. The collection of claim 66 in which the other substance is selected from antigens, drugs, vitamins, poisons, and pathogens.

71. The collection of claim 40 in which at least one fluorescent dye in the particles of each particle population is a lipid-soluble rhodamine comprising one of structures (I), (II), (III) and (IV).

72. The collection of claim 40 in which each particle population fluoresces when illuminated by the same excitation source.

73. The collection of claim 40 in which two or more fluorescent dyes fluoresce when irradiated with multiple excitation sources.

74. The collection of claim 40 in which at least one of the particle populations comprises a plurality of fluorescent dyes.

75. The collection of claim 40 in which the fluorescent dyes of the plurality are each, independently of one another, selected from lipid soluble rhodamine dye comprising structures (I), (II), (III) or (IV), acridine dyes, alizarene dyes, azo dyes, anthraquinine dyes, bodipy dyes, coumarin dyes, cyanine dyes, fluorescein dyes, lanthanide complexes, oxazine dyes, phenazathionium dyes, phenazoxonium dyes, porphyorin dyes, pyrene dyes, pyrilium dyes, perylene dyes, phenoxazine dyes, phenezine dyes, rhodol dyes and xanthene dyes.

76. The collection of claim 40 wherein the particles comprising the lipid soluble rhodamine additionally comprise an energy transfer dye.

77. A fluorescent polymeric material comprising a polymer and an energy transfer dye comprising two to four covalently attached dyes wherein one of said covalently attached dyes is a lipid soluble rhodamine dye that comprises one of the following “

core”

structures;

78. The fluorescent polymeric material of claim 77 wherein LK is selected from an ester, an amide, a sulfonamide, a hydrazine, an imine, a maleimide, a sulfide, a disulfide, a carbamate and a thiocarbamate.

79. The fluorescent polymeric material of claim 77 wherein all of the attached dyes are lipid soluble rhodamines.

80. A fluorescent polymeric material comprising a polymeric particle and a lipid soluble rhodamine.