Ammonia (NH3) has been identified as a potential hydrogen-carrier fuel with no carbon emissions. Non-intrusive in-situ NH3 diagnostic technique is of great interest. In this work, femtosecond two-photon laser-induced fluorescence (fs-TPLIF) was demonstrated in NH3/N-2 mixtures to achieve NH3 measurements. A femtosecond laser at 305 nm was used for two-photon excitation of NH3 to its excited state (X-C'), and the subsequent fluorescence at similar to 565 nm from transition C'-A was detected. In addition, a detection limit of 730 ppm was achieved in NH3 /N-2 mixtures. Furthermore, one-dimensional single-shot images of NH3 were obtained in both laminar and turbulent flow fields. This work is the first attempt of fs-TPLIF for polyatomic molecular gases measurements, and the obtained results indicate that fs-TPLIF could be a promising tool for NH3 measurements. (C) 2019 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.