Individual and population quantitative analyses of calcium flux in T-cells activated on functionalized material surfaces

We have developed a novel method for activating T-cells on material surfaces that enable individual and population-based analyses of intracellular calcium flux, as a quantitative measure of T-cell receptor engagement. Functionalized material surfaces were created using a plasma-polymerized foundation layer to immobilize stimulatory T-cell ligands, which could induce T-cell receptor-dependent calcium flux in naive T-cells. Real-time confocal microscopic detection and quantification of calcium flux using paired fluorescent ratiometric probes facilitated the tracking and analysis of response profiles of individual T-cells, as well as population analyses using a combination of individual T-cell events. This type of combined analysis cannot be achieved using traditional population-based flow cytometric approaches, and thus provides a logical step towards developing the capacity to assess the magnitude and quality of inherently heterogeneous effector T-cell responses to antigenic challenge.Susan N. Christo, Ghafar. T. Sarvestani, Stefani S. Griesser, Bryan R. Coad, Hans J. Griesser, Krasimir Vasilev, Michael P. Brown, Kerrilyn R. Diener and John D. Haybal