We evaluated the histologic features of the internal limiting membrane (ILM) of the retina removed during macular hole surgery without indocyanine green staining. Our investigation focused on the presence or absence of retinal structures adherent to the retinal surface of the ILM. Because only tiny retinal cellular fragments were observed especially in ILM folds, we conclude that conventional ILM peeling can be performed safely with a cleavage plane between the retinal surface of the ILM and Müller cell endfeet.