Objective To study the utility of fibrin glue on prevention of bleeding after endoscopic submucosal dissection (ESD). Methods Consecutive patients with gastrointestinal tumors who underwent ESD between July 2015 and June 2016 in the first Affiliated Hospital of Nanjing Medical University were enrolled in the study. The patients receiving ESD before December 31, 2015 were assigned into the fibrin glue group, and patients receiving ESD after December 31, 2015 were assigned into the control group. The treated group was sprayed with fibrin glue on wound post-ESD followed by routine hemostasis method, and the control group was given routine hemostasis method only. The bleeding rate after ESD, mean hospital stays and cost were compared between the two groups. Results The bleeding rate after ESD in the fibrin glue group was significantly lower than that in the control group [7.45% (12/161) VS 14.79% (25/169), P=0.035]. There was no significantly difference in the mean hospital stays (9.09±2.65 days VS 9.20±2.99 days, P=0.744) and mean cost (24 246±5 519 yuan VS 25 214±6 258 yuan, P=0.138) between the two groups. Multivariate analysis revealed that the use of fibrin glue was a protective factor for bleeding after ESD. Conclusion Fibrin glue is safe, effective and economical in prevention of bleeding after ESD.