Abstract: A multiphoton luminescence excitation microscopy utilizing a digital micromirror device (DMD), wherein the multiphoton luminescence excitation microscopy generates luminescence excitation light information according to excitation of a sample, comprising an excitation light source, a plurality of lenses, a digital micromirror device (DMD), an objective lens, a dichroic mirror and a light detector. The DMD is utilized to replace the conventional diffraction grating of luminescence excitation microscopy The DMD has advantages of easy accessibility, relatively low cost, successfully achieves temporal-focusing at the image formation plane, and the grating effect.