Hydrogel nanoparticles, poly(N-isopropylacrylamide-co-acrylic acid) (poly(NIPAm-co-AAc)), were synthesized by precipitation polymerization(32 ) and covalently functionalized with amino-containing dyes using amidation chemistry.(33 ) Zero-length cross-linking amidation strategies were performed in water or organic solvents on the basis of the hydrophilic/hydrophobic properties of the dyes. Amine-containing hydrogel nanoparticles, poly(NIPAm-co-AA), were created by precipitation polymerization and covalently functionalized with Cibacron Blue F3GA (CB) dye by nucleophilic substitution of the amine groups in the nanoparticles and a chloride atom in the CB dye. An outer shell containing vinylsulfonic acid (VSA) copolymer was created on the dye-functionalized particles by a second polymerization reaction as described below.