A high-resolution fluorescence image in which an afterimage is suppressed is obtained, even when a fluorescence detection interval is shortened. Provided is a scanning observation apparatus including a scanning unit that spatially scans pulsed excitation light emitted from a light source at prescribed time intervals on a specimen; a fluorescence detecting unit that detects fluorescence generated by exciting a fluorescent substance inside the specimen with the excitation light scanned by the scanning unit, in synchronization with the emission of the excitation light; and a fluorescence correcting unit that subtracts, from a fluorescence intensity detected by the fluorescence detecting unit, an afterimage fluorescence component calculated on the basis of time-sequential fluorescence detected by the fluorescence detecting unit prior thereto, at each scanning position, to correct the fluorescence intensity at the scanning position.