Prolonged hypotony-induced maculopathy is a serious complication of trabeculectomy with adjunctive mitomycin C. We performed trabeculectomies with intraoperative mitomycin C on 59 eyes of 52 consecutive patients. Exposure time to mitomycin C was five minutes in the first seven patients, two of whom had prolonged hypotony-induced maculopathy. One of these required surgical revision of the filtering procedure. Light and electron microscopic study of the excised, avascular bleb disclosed an irregular epithelium and largely acellular subepithelium of loosely arranged connective tissue. In the remaining 52 eyes, the exposure time to mitomycin C was titrated between two and five minutes according to each patient's risk for failure of filtration from excessive fibrosis. Four additional cases of prolonged hypotony-induced maculopathy occurred among these 52 cases (7.7%), all of which were in the lower risk groups that received two- or three-minute exposure to mitomycin C. Four procedures failed, requiring further glaucoma surgery, and all of the patients were in the higher risk groups, receiving three- to five-minute exposures. Our titration of the exposure time to mitomycin C may have reduced, but did not eliminate, the risk fo prolonged hypotony-induced maculopathy, and further study is needed to establish the optimum protocol for the use of this drug as an adjunct to trabeculectomy.