Two-photon fluorescence (TPF) of olive oils is discovered and observed experimentally for the first time. Variations of the single-photon fluorescence (SPF) and TPF with the excitation wavelength are investigated for four different olive oils. The results show that fluorescence of the cosmetic olive oils (COO) is very weak and exhibits only one spectral peak around 490nm. While for the ordinary edible oils (OEO) whether they are during their shelf life or not, their fluorescence spectra may exhibit multiple peak structures. The short-term natural expiration only slightly weakens TPF of OEO. Moreover, the excitation wavelength affects the OEO spectra considerably in terms of the spectral peak number, the spectral peak position, and spectral shapes. When the excitation wavelength decreases from 700nm, the whole TPF of the OEO also decreases. Relatively, however, the short wave band will decrease and disappear more quickly. While for the SPF, the long wave band will decrease and disappear first. The optimal excitation wavelengths to make the TPF strongest are around 700nm and 640nm for OEOs and COO, respectively. And effects of temperature on SPF and TPF of extra virgin olive oil are also explored. This work may be of significance for its potential applications in TPF detection and two-photon laser.