In this study, we evaluated the influence of retrieval algorithms and sensor characteristics, such as spectral resolution (SR) and signal to noise ratio (SNR), on the retrieval accuracy of fluorescence signal (Fs). Here Fs was retrieved by four commonly used retrieval methods, namely the original Fraunhofer Line Discriminator method (FLD), the 3 bands FLD (3FLD), the improved FLD (iFLD) and the spectral fitting method (SFM). Fs was retrieved in the oxygen A band centered at around 761nm (O2-A). We analyzed the impact of sensor characteristics on four retrieval methods based on simulated data which were generated by the model SCOPE (Soil Canopy Observation, Photochemistry and Energy fluxes), and obtained consistent conclusions when compared with experimental data. Results presented in this study indicate that both retrieval algorithms and sensor characteristics affect the retrieval accuracy of Fs. When applied to the actual measurement, we should choose the instrument with higher performance and adopt appropriate retrieval method according to measuring instruments and conditions.