A chemical signal generator for resolving temporal dynamics of single cells.

To investigate rapid cell signaling, analytical methods are required that can generate repeatable chemical signals for stimulating live cells with high temporal resolution. Here, we present a chemical signal generator based on hydrodynamic gating, permitting flexible stimulation of single adherent cells with a temporal resolution of 20 ms. Studies of adenosine triphosphate (ATP)-induced calcium signaling in HeLa cells were demonstrated using this developed method. Consecutive treatment of the cells with ATP pulses of 20 or 1 s led to an increase of latency, which might be another indicator of receptor desensitization in addition to the decrease in the amplitude of calcium spikes. With increasing duration of ATP pulses from milliseconds to a few seconds, the cellular responses transitioned from single calcium spikes to calcium oscillation gradually. We expected this method to open up a new avenue for potential investigation of rapid cell signaling.