A fluorescence detecting apparatus includes: an excitation light emitter for emitting excitation light exciting a fluorescent material onto a measurement object; an illumination light emitter for emitting illumination light onto the measurement object; an image pickup device for sensing light from the measurement object; and a controller for controlling the image pickup device, the excitation light emitter, and the illumination light emitter. The excitation light is emitted from the excitation light emitter onto the measurement object to define an image pickup condition of the image pickup device, based on an image pickup result to be obtained by sensing light from the measurement object by the image pickup device. A light amount of the illumination light to be emitted from the illumination light emitter is set depending on the image pickup condition. The excitation light, and the illumination light of the set light amount are simultaneously emitted from the excitation light emitter and the illumination light emitter, to cause the image pickup device to sense light from the measurement object.