We report a two-photon fluorescent probe which shows a strong two photon excited fluorescence enhancement in response to Zn(2+), easy loading into the cells, Golgi-localizing ability, low cytotoxicity, and high photostability. Two-photon microscopy imaging revealed that this probe allows for real-time monitoring of the changes in Golgi Zn(2+) as well as their 3D distributions in live cells and tissues.