Fluorescence-Based Zinc Ion Sensor for Zinc Ion Release from Pancreatic Cells

Abstract

In this paper, we describe the synthesis and characterization of analytical properties of fluorescence-based zinc ion-sensing glass slides and their application in monitoring zinc ion release from beta pancreatic cells in cell cultures. To fabricate the sensors, the zinc ion indicator ZnAF-2 {6-[N-[N',N'-bis(2-pyridinylmethyl)-2-aminoethyl]amino-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one} was modified to include a sufficiently long linking aliphatic chain with a terminal carboxyl functional group. The recently synthesized ZnAF-2 zinc ion indicator provided high zinc ion selectivity in physiological solutions containing millimolar levels of calcium and other possible interfering cations. The carboxyl-modified ZnAF-2 was conjugated to the activated surface of glass slides, which then served as zinc ion sensors. It was possible to grow pancreatic cells directly on the zinc-sensing glass slide or on a membrane placed on these glass slides. The sensors were used to monitor zinc ion release events from glucose-stimulated pancreatic cells. The study showed that the zinc ion sensors responded effectively to the release of zinc ions from pancreatic cells at the nanomolar level with high selectivity and rapid subsecond response time.