The invention provides a method for making a tissue chip based on one-step molding. The method is characterized in that the tissue chip is made through one-step molding without preparing a donor which is paraffin or an acceptor which is a paraffin tissue block. The method concretely comprises the following steps: 1, taking a mold frame, putting a tissue in the embedding tube of the mold frame in a required size and a required tangent plane, dumping paraffin for embedding, and allowing the paraffin to stand for a period of time to carry out polymerization molding of the paraffin; and 2, taking a paraffin-embedded frame, placing the mold frame having the embedding structure obtained in step 1 in the embedding frame on flat glass, dumping molten paraffin into the paraffin-embedded frame, allowing the paraffin in the porous tubes of the mold frame and the paraffin in the embedded frame to melt, and removing the embedded frame after the melting to complete the tissue embedding. According to the invention, the tissue positions and the embedding accuracy are strictly controlled, and no burrs or discontinuous cracks appear during punching, so the method has the advantages of cheapness, convenience, high efficiency, time saving and labor saving, and can be popularized to each pathology department and relevant research units.