On a substrate 41 holding a sample to be detected, a dielectric multilayer 42 is disposed which reflects excitation light e1 supplied from above the substrate 41 and transmits fluorescence f1 emitted from the sample, and the excitation light e1 is reflected at the dielectric multilayer 42 while the transmitted fluorescence f1 is detected by a light receiving unit 44, thereby providing a fluorescence measurement apparatus which can resolve a problem of reduction in detection sensitivity due to autofluorescence from the substrate or leakage of the excitation light from a light receiving filter, and which can detect the sample with high sensitivity.