Visualization and quantification of endoplasmic reticulum Ca2+ in renal cells using confocal microscopy and Fluo5F

► A new method of monitoring ER Ca2+ directly in live cells. ► Confocal microscopy and low affinity Ca2+ indicator, Fluo5F, reveals ER Ca2+ morphology. ► Fluo5F fluorescence shows no co-localization with mitochondria or cytosol. ► Stimulation of ER Ca2+ release reveal spatiotemporal differences in ER Ca2+ release dynamics.