Provided is a microscope optical system in which the occurrence of flare due to unnecessary-order diffracted light exited from a diffractive optical element is suppressed. A microscope objective lens MS is configured by including an objective lens OL which has a diffractive optical element GD and converts light from an object into a substantially parallel light flux, and a second objective lens IL which forms an image of the object by focusing the substantially parallel light flux from the objective lens OL, and is configured such that, in case where an m-th order of diffracted light from the diffractive optical element GD is used for the image formation, the following expression is satisfied: |θ|>tan−1(0.06/D) when the light of a maximum NA emitted from the object located on an optical axis enters the diffractive optical element.