Precision fluorescently dyed polymeric microparticles and methods of making and using same

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:

• 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 a series of solutions, each comprising at least two fluorescent dyes in a solvent, wherein each of the solutions in the series 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 emits a different targeted fluorescence intensity from the at least one of the at least two fluorescent dyes upon excitation, and wherein the different targeted fluorescence intensities have been used to determine the different concentrations of the at least two fluorescent dyes included within each of the solutions.

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:

• 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 a series of solutions, each comprising at least two fluorochromes in a solvent, wherein each of the solutions in the series 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 emits a different targeted fluorescence intensity from the at least two fluorochromes upon excitation, and wherein the different targeted fluorescence intensities have been used to determine the different concentrations of the at least two fluorochromes included within each of the solutions.