The investigation was undertaken to study whether femtosecond laser ablation and microscopy might be used in the internal retinal borderline membrane. Ablation of internal limiting membrane preparations removed using or not using indocyanine green was made by a low-energy femtosecond laser. Examination of the preparations by laser and electron microscopy revealed precision laser cuts of the internal retinal borderline membrane. The use of indocyanine green during laser ablation reduced laser irradiation parameters as compared to the dye not being applied. Low-energy femtosecond lasers enable precision contactless ablation of the internal borderline membrane to be carried out without collateral damage to the adjacent tissue. The parameters of laser impulses, particularly low ones used in the ablation of indocyanine green-stained preparations, prove the photosensitizing effect of the dye.