The present invention discloses a method for non-fluorescence higher harmonic generation ground state depletion super-resolution microscopy, it includes the following steps: providing an organic material unit, focusing excitation light and ground state depletion light, generating a higher harmonic signal, performing ground state depletion and performing microscopic imaging. With the implementation of the present invention, the stimulated electrons of the organic material remains majorly on the singlet (S1) state or the triplet (T1) state, instead of the ground (S0) state, to provide modulation of the spatial distribution of the non-fluorescence signal, and make STED microscopy applicable to non-fluorescence signals to promote the resolution of the images.