Purpose To examine three layers of membranes detected by triamcinolone acetonide (TA) and indocyanine green (ICG) during surgery for macular hole and retinal detachment (MHRD) as a result of high myopia. Design Histologic study. Methods We excised three layers of membranes visualized with TA and ICG that were tightly adhering to the retinal surface during vitrectomy and examined them by transmission electron microscopy. Results The first membrane was made of acellular collagen fibers, suggestive of vitreous, and the second was comprised of parts of fibroblast-like cells and collagen fibrils, suggesting a proliferative epiretinal membrane. The third membrane was an internal limiting membrane. Conclusions Multiple components of epiretinal tissue could be delaminated with the assistance of TA and ICG during surgery for myopic MHRD.