In this study, we established a new fluorescent indicator platform. The responsive element consists of poly(N-isopropylacrylamide) nanospheres that include small percentages of fluorescein and a ligand, anilinodiacetate (phenylIDA). Nanosphere diameters were determined to be in the range from 50 to 90 nm by scanning electron microscopy. They were entrapped in a polyacrylamide gel to prevent nanosphere aggregation. At pH 6, the ligand is negatively charged in the absence of metal ions. Charge-charge repulsion causes the nanosphere to swell. Dynamic light scattering measurements show that these nanospheres do not shrink and aggregate at high temperature. Cu(II) binding neutralizes the charge causing the particles to shrink. This brings fluoresceins closer together, increasing the degree of self-quenching. The intensity decreases by 30% as Cu(II) concentration increases. To rule out the possibility that the observed decrease in intensity was due to Cu(II) quenching of fluorescence, we also added Zn(II) and observed a decrease in intensity. This approach can be adapted to sense different metal ions and different concentrations of Cu(II) by changing the ligand.
