The invention provides a spider intestinal tract dissection method, and belongs to the field of arthropod anatomy. According to the method, dissection is started from back, heart and feces sac are taken as a breakthrough, and the intestinal tract position can be quickly and accurately found; meanwhile, the integrality of the intestinal tract can be guaranteed to the maximum extent. The whole dissection process only needs 5-10 minutes, the dissection time is greatly shortened, the pollution and the waste of spider materials are reduced, a favorable experimental material basis is provided for the follow-up researches of pest prevention and control, spider intestinal tract microorganism and the like.