Encapsulation of discrete quanta of fluorescent particles

• US 6,905,766 B2
• Filed: 01/31/2003
• Issued: 06/14/2005
• Est. Priority Date: 08/17/1999
• Status: Expired due to Term

• Current Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Original Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Inventors: Chandler, Don J.
• Primary Examiner(s): Acquah, Samuel A.
• Application Number:
  US10/355,073
• Publication Number:
  US 20030132538A1
• Time in Patent Office: 865 Days
• Field of Search: 264/4.1, 264/4.3, 264/4.33, 264/4.4, 264/4.7, 428/402.2, 428/402.21, 428/403, 428/690
• US Class Current: 428/402.2
• CPC Class Codes: B01J 13/046 combined with gelification or coagulation
• CPC Class Codes: B01J 13/18 In situ polymerisation with all reactants being present in the same phase
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: G01N 2015/1014 Constitution of reference particles
• CPC Class Codes: G01N 2333/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: G01N 2400/26 Cellulose
• CPC Class Codes: G01N 2400/28 Chitin, chitosan
• CPC Class Codes: G01N 33/587 Nanoparticles
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]
• CPC Class Codes: Y10T 428/2985 Solid-walled microcapsule from synthetic polymer
• CPC Class Codes: Y10T 428/2991 Coated

Abstract

The present invention provides novel encapsulation compositions and methods. In particular, the invention relates to fluorescent capsule compositions, which consists of a layer of a polymer shell enclosing one or more fluorescent materials such as fluorescent microspheres and which are capable of emitting at least two distinct fluorescent signals. Also provided are methods for their preparation. The compositions and methods of this invention are useful in a variety of applications, including preparation of multiplexed arrays for industrial, chemical, immunological, and genetic manipulation and analysis especially as related but not limited to flow cytometry.

Claims

1. A composition comprising at least two types of discrete fluorescent materials, each capable of emitting a distinct fluorescent light signal, said discrete fluorescent materials encapsulated by a shell barrier.

2. The composition of claim 1 wherein the discrete fluorescent materials comprise microspheres or nanospheres.

3. The composition of claim 1 wherein the discrete fluorescent materials comprise powders, crystals, rods, fibers, liquid, or a combination thereof.

4. The composition of claim 1 wherein the shell barrier comprises a layer of a polymerizable material.

5. The composition of claim 1 wherein the composition exhibits a spherical shape or a granular shape.

6. The composition of claim 1 whereby said composition has a diameter of about 1 nanometer to about 10 millimeters.

7. A fluorescence encapsulation device comprising an inner zone delimited by a capsule shell that inhibits the leakage of fluorescence emitting materials, wherein the capsule shell comprises a hydrophilic or hydrophobic polymer material, wherein the inner zone comprise two or more fluorescent light emitting materials each capable of emitting at least one distinct fluorecent signal.

8. A method of making a matrix, which emits at least two distinct light signals, the method comprising the steps of:

a) mixing in a matrix-forming material at least two light emitting materials at a desired ratio; and

b) polymerizing the mixture obtained at step (a) to form the matrix, where said polymerizing will deter leakage of the at least two light emitting materials out of the matrix.

16. The method of claim 8, wherein an outer surface of the matrix consitutes a barrier encapsulating the at least two light emitting materials.

17. The method of claim 8, wherein said mixing comprises dispersing the at least two light emitting materials in the matrix-forming material.

18. The method of claim 8, wherein the at least two light emitting materials comprises soluble or insoluble fluorescent materials.

19. The method of claim 8, wherein the at least two light emitting materials are presented as spheres, powders, crystals, rods, fibers, liquids, or a combination thereof.

20. The method of claim 8, wherein said polymerizing comprising crosslinking the mixture to form solid particles of the matrix.

21. The method of claim 8, further comprising mixing in the matrix-forming material additional components that will further deter the leakage of the at least two light emitting materials out of the matrix.

22. The method of claim 8, wherein the at least two distinct light signals have precisely controlled intensities.

9. A process for encapsulating two or more fluorescent materials, each capable of emitting at least one distinct fluorescent signal, in a shell comprising:

a) dispersing or dissolving said two or more fluorescent materials in a matrix-forming material;

b) adding a shell material or precursor thereof to the two or more fluorescent materials in the matrix-forming materials; and

c) polymerizing the shell material or its precursor to a polymerized shell which encapsulates the two or more fluorescent materials.

10. A process for encapsulating fluorescent materials in capsule shell, wherein the capsule shell emits two or more distinctive fluorescent signals, the process comprising:

a) preparing an encapsulation composition by dispering or dissolving said fluorescent material in a matrix-forming material,

b) emulsifying the encapsulation composition in a solution of gelatin,

c) mixing at desired rate the obtained solution to allow said encapsulation composition to break up into droplets of desired size,

d) adding a solution of gum arabic,

e) thermally softening the obtained solution while adding warm water,

f) bringing the pH of the solution to about 4.0 and 4,4,

g) cooling the solution and adding a cross-linking agent to form the capsule shell,

h) bringing the pH to about 9, and;

i) curing the capsule shell by raising the temperature to about 50°

C.

11. A composition comprising (i) two or more substances, each substance capable of emitting a distinctive fluorescent signal, and (ii) a shell barrier encapsulating said two or more substances.

12. The composition of claim 11 in which said fluorescent signal is distinctive by way of its wavelenght.

13. The composition of claim 11 in which said flourescent signal is distinctive by way of its intensity.

14. The composition of claim 11 in which said flourescent signal is distinctive by way of its wavelenght, intensity, or both.

15. The composition of claim 11 in which said each substance comprises one or more particles each stained with a fluorescent dye.

23. A matrix, comprising:

a polymerized matrix-forming material; and

at least two light emitting materials contained within the polymerized matrix-forming material, wherein polymerization of the matrix-forming matrial deters leakage of the at least two light emitting materials out of the matrix.

24. The matrix of claim 23, wherein an outer surface of the matrix constitutes a barrier encapsulating the at least two light emitting materials.

25. The matrix of claim 23, wherein the at least two light emitting materials comprise soluble or insoluble fluorescent materials.

26. The matrix of claim 23, wherein the at least two light emitting materials are presented as spheres, powders, crystals, rods, fibers, liquids, or a combination thereof.

27. The matrix of claim 23, wherein the polymerized matrix-forming material comprises a crosslinked matrix-forming material that forms solid particles of the matrix.

28. The matrix of claim 23, further comprising additional components contained within the polymerized material-forming material that will further deter the leakage of the at least two light emitting materials out of the matrix.

29. The matrix of claim 23, wherein the at least two light emitting materials emit distinct light signals having precisely controlled intensities.