Measurement of Calcium Signaling in Beta-Cell Lines Using Epifluorescence and Confocal Microscopy

One of the main consequences of glucose action on pancreatic β-cells is the stimulation of Ca²⁺ entry as well as Ca²⁺ release from intracellular compartments. Therefore, one of the cornerstones of any diabetes research laboratory should be the ability to measure changes in intracellular calcium concentrations within pancreatic β-cells. There are a variety of methods available for the measurement of intracellular calcium, from multiple regions of interest (ROIs) within single cells to observe oscillating calcium, to population-based 96-well applications to enable high-throughput screening of the effects of novel agonists. These methods allow calcium signaling to be observed in a single cellular assay to look at oscillations at a cellular level, to view a population response, and to enable high-throughput assays where the mean reflects a single cell response.