Fluorescent polymeric materials containing lipid soluble rhodamine dyes

• US 8,618,161 B2
• Filed: 02/14/2011
• Issued: 12/31/2013
• 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: Lam, Joe Y. L., Menchen, Steven M., Zou, Ruiming, Benson, Scott C.
• Primary Examiner(s): Chu, Yong
• Assistant Examiner(s): Cheng, Karen
• Application Number:
  US13/026,690
• Publication Number:
  US 20110226994A1
• Time in Patent Office: 1,051 Days
• Field of Search: 514/453, 549/224, 549/382
• US Class Current: 514/453
• 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 polymeric particle and one or more lipid soluble rhodamine dyes, comprising the following core structure:

wherein R¹¹ and R¹⁵ are each H or are each F or Cl; R¹², R¹³, and R¹⁴ are each H or are each F or Cl; and wherein at least one of R^3′, R^3″, R^6′ and R^6′ is a (C4-C20) alkyl. 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 polymeric particle and at least one lipid-soluble rhodamine dye comprising the following core structure:

2. The fluorescent polymeric material of claim 1, wherein the polymeric particle is composed of a particle formed by polymerizing one or more of the following monomers:

substituted or unsubstituted styrenes, acrylate, alkyl acrylates, methacrylates, 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 a crosslinking agent.

3. The fluorescent polymeric material of claim 1, wherein the polymeric particle is composed of a hydrophobic copolymer of styrene, acrylic acid and a polyvinyl crosslinking agent.

4. The fluorescent polymeric material of claim 1, wherein the polymeric particle is in the form of a microparticle having a diameter in the range of about 0.01 to 1000 micrometers.

5. The fluorescent polymeric material of claim 1 wherein the polymeric particle is activated with one or more groups that comprise a reactive functionality.

6. The fluorescent polymeric material of claim 5, wherein the reactive functionality is an electrophile selected from the group consisting of pentafluorphenyl ester, NHS-ester, acrylamides, acyl azides, acyl halides, aldehydes or ketones, alkyl halides, alkyl sulfonates, anhydrides, aryl halides, aziridines, bonorates, carboxylic acids, carbodiimides, diazoalkenes, epoxides, haloacetamides, halotriazines, imido esters, isocyanates, isothiocyanates, maleimides, phosphoramidites, silyl halides, sulfonate esters and sulfonyl halides.

7. The fluorescent polymeric material of claim 5, wherein the reactive functionality is a nucleophile selected from the group consisting of amines/anilines, alcohols/phenols, thiols, hydrazines and hydroxylamines.

8. The fluorescent polymeric material of claim 1, further comprising another substance that is absorbed into, adsorbed onto or into, or covalently attached to, the polymeric particle.

9. The fluorescent polymeric material of claim 8, wherein the another substance is selected from the group consisting of a biomolecule and a probe for a biomolecule.

10. The fluorescent polymeric material of claim 1, further comprising from one to nine additional fluorescent dye types non-covalently incorporated into the polymeric particle.

11. The fluorescent polymeric material of claim 10, wherein each of the additional fluorescent dye types is, independently of the others, selected from the group consisting of:

a lipid-soluble rhodamine dye according to structural formula (IV), an additional lipid-soluble rhodamine dye and lipid-soluble form of;

an acridine dye, an alizarine dye, an azo dye, an anthraquinine dye, a BODIPY dye, a coumarin dye, a cyanine dye, a fluorescein dye, a lanthanide complex, an oxazine dye, a phenazathionium dye, a phenazoxonium dye, a porphyrin dye, a pyrene dye, a pyrilium dye, a perylene dye, a phenoxazine dye, a phenezine dye, a rhodol dye and a xanthene dye.

12. The fluorescent polymeric material of claim 10, wherein all of the fluorescent dyes fluoresce when irradiated with the same excitation source.

13. The fluorescent polymeric material of claim 10, wherein two or more of the fluorescent dyes fluoresce when irradiated with multiple excitation sources.

14. The fluorescent polymeric material of claim 10, wherein at least one of the fluorescent dyes is an energy transfer dye.

15. A collection of fluorescent polymeric particles, each of which has a unique, spectrally resolvable fluorescence emissions spectrum, wherein at least one of the polymeric particles is a polymeric material according to claim 1.