A technique for evaluating calcium flux in primary airway tissue at single cell resolution

ABSTRACT Calcium signaling is critical in a multitude of biological processes, reinforcing the need to develop methods to study calcium flux in primary tissue. The study of live calcium mobilization into the cytosol is commonly studied by intracellular calcium dyes. However, the study of calcium mobilization at a single-cell resolution in differentiated, primary human tissue remains challenging. Here we present methodology to study calcium flux at single cell resolution in situ on primary tissue seeded on trans wells. As well, we developed a machine learning software that simplifies and streamlines analysis. SUMMARY Live intracellular calcium dyes enable our group to study cell type specific responses as fluorescent readouts in real time. Utilizing microscopic techniques, we devised methods to study calcium flux in individual cells on complex, differentiated primary tissues seeded on trans well inserts.