Precision fluorescently dyed particles and methods of making and using same
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Abstract
An improved method of making a series of bead or microsphere or particle populations characterized by subtle variation in a proportion or ratio of at least two fluorescent dyes distributed within a single bead of each population is provided. These beads, when excited by a single excitation light source are capable of giving off several fluorescent signals simultaneously. A set containing as many as 64 distinct populations of multicolored, fluorescent beads is provided and when combined with analytical reagents bound to the surface of such beads is extremely useful for multiplexed analysis of a plurality of analytes in a single sample. Thus, methods of staining polymeric particles, the particles themselves, and methods of using such particles are claimed.
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Citations
96 Claims
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1-40. -40. (canceled)
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41. A population of polymeric microspheres dyed with two or more fluorescent dyes, obtained by a method comprising:
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treating polymeric microspheres by adding a polar organic solvent to an aqueous solution containing the polymeric microspheres to produce a mixture; and
adding at least two fluorescent dyes to the mixture such that the polymeric microspheres are stained with the at least two fluorescent dyes, wherein the at least two fluorescent dyes are selected such that on isolation and excitation of the polymeric microspheres, a distinct fluorescence signal is emitted from each dye, and wherein the distinct fluorescence signals have an intensity proportional to the amount of the dye in the polymeric microspheres. - View Dependent Claims (42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54)
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62. A set of distinct populations of polymeric microspheres dyed with at least two fluorescent dyes, wherein the set of distinct populations of polymeric microspheres is dyed using a method comprising:
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treating separate populations of polymeric microspheres by adding a polar organic solvent to aqueous solutions containing the separate populations of polymeric microspheres to produce different mixtures;
preparing solutions of at least two fluorescent dyes, wherein each of the solutions has a different concentration of at least one of the at least two fluorescent dyes; and
adding said each of the solutions to one of the different mixtures to provide the set of distinct populations of polymeric microspheres, wherein each of the distinct populations of polymeric microspheres has a different fluorescence intensity from the at least one of the at least two fluorescent dyes. - View Dependent Claims (63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82)
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83. A population of polymeric microspheres dyed with two or more fluorochromes, obtained by a method comprising:
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treating polymeric microspheres by adding a polar organic solvent to an aqueous solution containing the polymeric microspheres to produce a mixture; and
adding at least two fluorochromes to the mixture such that the polymeric microspheres are stained with the at least two fluorochromes, wherein the at least two fluorochromes are selected such that on isolation and excitation of the polymeric microspheres, a distinct fluorescence signal is emitted from each fluorochrome, and wherein the distinct fluorescence signals have an intensity proportional to the amount of the fluorochrome in the polymeric microspheres. - View Dependent Claims (84, 85)
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86. A set of distinct populations of polymeric microspheres dyed with at least two fluorochromes, wherein the set of distinct populations of polymeric microspheres is dyed using a method comprising:
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treating separate populations of polymeric microspheres by adding a polar organic solvent to aqueous solutions containing the separate populations of polymeric microspheres to produce different mixtures;
preparing solutions of at least two fluorochromes, wherein each of the solutions has a different concentration of at least one of the at least two fluorochromes; and
adding said each of the solutions to one of the different mixtures to provide the set of distinct populations of polymeric microspheres, wherein each of the distinct populations of polymeric microspheres has a different fluorescence intensity from that at least one of the at least two fluorochromes. - View Dependent Claims (87, 88, 89, 90, 91, 92, 93, 94, 95, 96)
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Specification