Choroidal melanoma is the most frequent form of primary neoplasia among malignant ocular tumors. Since it is presumed that metastasis often occurs before the primary tumor is first diagnosed, early detection is exigent. The aim of the studies described in this report was to develop an objective, noninvasive method for the diagnosis of choroidal melanoma. The underlying new principle of fluorescence excitation is presented. This is based on the observation that melanin, due to its unique absorption characteristics, is selectively excited into fluorescence via stepwise absorption of two photons of a femtosecond laser emitting at 800 nm. In the experiment described, the fluorescence of excised tissue from healthy choroidal pigment epithelium was compared to that of excised choroidal melanoma. The fluorescence of choroidal melanomas exhibited a more reddish appearance and less intensity than that of healthy tissue. This implies that the configuration of melanin apparently changes during the process of malignant degeneration. The method described here could thus serve as an evidentiary objective diagnostic technique before initiating treatment for choroidal melanomas.