Abstract: The present invention is related to a three-dimensional graphene oxide microstructure and making method of thereof. First, a photoreactive agent is added into a graphene oxide solution, wherein the photoreactive agent is a photoreactivator in a nonlinear optical method. Then, the photoreactivator in the graphene oxide solution is activated by a beam emitted from an excitation module to produce singlet oxygen with high activity. Finally, the graphene oxide is activated by the singlet oxygen for an unpaired electron of the graphene oxide covalently bonding with another graphene oxide to form a three-dimensional graphene oxide microstructure. Therefore, two-dimensional graphene oxides are efficiently cross-linked with each other to form a three-dimensional graphene oxide microstructure by a nonlinear optical technique of an ultrafast laser system so as to apply to the development of all electronic and optical components.