This fluorescence observation method is a method for observing a living organism into which a fluorescent dye is injected. The fluorescence observation method is provided with: a step for irradiating the living organism with excitation light including a wavelength for exciting the fluorescent dye using a light irradiation means; a step for acquiring a first fluorescence image of the living organism using an image acquisition means, the first fluorescence image being generated by the irradiation with the excitation light; a step for specifying an object to be observed in the living organism on the basis of the first fluorescence image; a step for acquiring a second fluorescence image of the object to be observed using the image acquisition means, the second fluorescence image being generated by the irradiation with the excitation light; and a step for specifying a linear fluorescence pattern appearing in the second fluorescence image.本發明係一種螢光觀察方法,其係用於觀察經注入螢光染料之生物體者,該方法具備如下步驟:利用光照射機構,朝生物體照射包含激發螢光染料之波長之激發光;利用圖像獲取機構,獲取藉由激發光之照射而產生的生物體之第1螢光圖像;基於第1螢光圖像而特定生物體中之觀察對象;利用圖像獲取機構,獲取藉由激發光之照射而產生的觀察對象之第2螢光圖像;及特定第2螢光圖像中顯現之線狀螢光圖案。