Precision fluorescently dyed particles and methods of making and using same
First Claim
1. A method of staining polymeric microspheres with two or more fluorescent dyes comprising:
- (a) combining at least two fluorescent dyes in a solvent mixture comprising at least one organic solvent in which the at least two fluorescent dyes are soluble and at least one alcoholic solvent in which the at least two fluorescent dyes are less soluble, to provide a solution of mixed dyes, which has the capacity to swell at least partially but not dissolve a plurality of polymeric microspheres brought into contact with said solution;
(b) contacting a plurality of polymeric microspheres with said solution for a period of time sufficient to provide uniform staining of substantially all of the members of said plurality of polymeric microspheres with the at least two fluorescent dyes, said at least two fluorescent dyes being selected such that on isolation an excitation of said dyed plurality of polymeric microspheres, a distinct fluorescence signal is emitted from each dye, the intensity of which emitted signal is proportional to the amount of said dye in said dyed plurality of polymeric microspheres;
(c) preparing a series of said solutions having differing desired concentrations of said at least two fluorescent dyes; and
(d) contacting separate populations of a plurality of polymeric microspheres with said series of said solutions to provide multiple distinct populations or subsets of a plurality of polymeric microspheres, each distinct population or subset having a differing desired fluorescence intensity of said at least two fluorescent dyes.
1 Assignment
0 Petitions
Accused Products
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.
-
Citations
18 Claims
-
1. A method of staining polymeric microspheres with two or more fluorescent dyes comprising:
-
(a) combining at least two fluorescent dyes in a solvent mixture comprising at least one organic solvent in which the at least two fluorescent dyes are soluble and at least one alcoholic solvent in which the at least two fluorescent dyes are less soluble, to provide a solution of mixed dyes, which has the capacity to swell at least partially but not dissolve a plurality of polymeric microspheres brought into contact with said solution;
(b) contacting a plurality of polymeric microspheres with said solution for a period of time sufficient to provide uniform staining of substantially all of the members of said plurality of polymeric microspheres with the at least two fluorescent dyes, said at least two fluorescent dyes being selected such that on isolation an excitation of said dyed plurality of polymeric microspheres, a distinct fluorescence signal is emitted from each dye, the intensity of which emitted signal is proportional to the amount of said dye in said dyed plurality of polymeric microspheres;
(c) preparing a series of said solutions having differing desired concentrations of said at least two fluorescent dyes; and
(d) contacting separate populations of a plurality of polymeric microspheres with said series of said solutions to provide multiple distinct populations or subsets of a plurality of polymeric microspheres, each distinct population or subset having a differing desired fluorescence intensity of said at least two fluorescent dyes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
-
Specification