Fluorescence observation apparatus of the present invention is emitted for fluorescent substance and a plurality of light sources capable of emitting section, a plurality of pump lights and the plurality of pump lights for exciting each of the plurality of fluorescent materials, and reference light I respectively generates an image signal in accordance with an imaging unit that captures a plurality of fluorescent emitted by, and reflected light of the reference light, a plurality of fluorescent taken by the imaging unit and to the reflected light of the reference light image to be output is assigned respectively to the color channels of the plurality, and a reference light image related fluorescence images according to the image generator, the image signal corresponding to the plurality of fluorescent, an image signal corresponding to the reflected light of the reference light I have, a processing unit.本発明の蛍光観察装置は、複数の蛍光物質をそれぞれ励起させるための複数の励起光と、参照光と、を出射可能な光源部と、複数の励起光を複数の蛍光物質に対して出射することにより発せられる複数の蛍光と、参照光の反射光と、を撮像する撮像部と、撮像部により撮像された複数の蛍光と、参照光の反射光と、に応じた画像信号をそれぞれ生成する画像生成部と、複数の蛍光に応じた画像信号に係る複数の蛍光画像と、参照光の反射光に応じた画像信号に係る参照光画像と、を複数の色チャンネルにそれぞれ割り当てて出力する画像処理部と、を有する。
