There is provided a fluorescence observation apparatus, including: a lighting section including a light source emitting illumination light and excitation light; a fluorescence imaging section picking up fluorescence generated on a subject and acquiring a fluorescence image; a returned light imaging section picking up returned light returned from the subject and acquiring a returned light image; a preprocessing section multiplying at least either the fluorescence image or the returned light image by a coefficient in which a distance property of fluorescence intensity and a distance property of returned light intensity, which are acquired in advance with respect to a standard sample, come to be in a relation directly proportional to each other and generating a fluorescence image for correction and a returned light image for correction; and a fluorescence image correction section dividing the fluorescence image for correction by the returned light image for correction.