The invention relates to an aqueous fluorescent invisible stitching plaster comprising the following materials by weight part: 10-20 parts of thickening agent, 0.1-0.9 part of decomposition agent, 0.5-1.5 parts of distiller liquor and 65-80 parts of purified water. A preparation method of the aqueous fluorescent invisible stitching plaster comprises the following steps: infusing the thickening agent, the decomposition agent and the distiller liquor into the purified water for 2-3 hours and then pouring into a stirrer to stir for 1.5-2 hours, and then adding a proper amount of fluorescent powder after cooling for barreling and packaging. The invention has the advantages that: 1. the aqueous fluorescent invisible stitching plaster can ensure that a printed white mark is invisible under natural light compared with the traditional white and grey stitching plaster, thereby avoiding the trouble of cleaning the vamp and saving the labour cost for cleaning; 2. the printing output can be increased by about 1/3 to 1/2, a halftone can be prevented from being blocked and is hard to scrap, printing is easier for workers, the output and the quality are more stable; and 3. the aqueous fluorescentinvisible stitching plaster can save much illumination power charge.