A micro-fluidic chip and an operation method thereof are disclosed. The micro-fluidic chip includes a detection base plate including a non-paper based substrate and an electrode layer on the substrate; and a paper-based chip laminated on the electrode layer and including a hydrophilic substrate, wherein the paper-based chip also includes a fluid passage on the hydrophilic substrate and a reactionzone communicated with the fluid passage, with the reaction zone being arranged corresponding to the electrode layer. In the micro-fluidic chip, the electrode layer is integrated on the non-paper based substrate so that the detection base plate can be repeatedly used, waste of noble metal electrode materials is avoided, the consumption cost is reduced, and environment pollution is avoided. The hydrophilic substrate is adopted as the substrate of the fluid passage and the reaction zone, and capillarity reaction of the hydrophilic substrate directs fluid to flow so that formation of a fluid passage with a complex etching process, preparation of a micro pump, and the like are not needed, thus greatly simplifying the manufacturing process.