To view this video please enable JavaScript, and consider upgrading to a web browser that supports HTML5 video Video 1 Typical TIRFM recording of Ca2+ puffs from a field of HEK293 cell stimulatedwith carbachol (10 μM) after ~20 s. The video, showing unprocessed images, was captured and replayed at 25 frames/s (fps). Scale bar = 10 μm. Endogenous signalling pathways and caged IP3 evoke Ca2+ puffs at the same abundant immobile intracellular sites Michael V. Keebler, and Colin W. Taylor J Cell Sci 2017. 130:3728-3739; doi: 10.1242/jcs.208520 Download Original Video