A manufacturing method of antireflection substrate structure includes: providing a silicon wafer having a first rough surface; forming an antireflection optical film on the silicon wafer, wherein the antireflection optical film conformally overlays the first rough surface; performing a surface treatment on the antireflection optical film so that the antireflection optical film has a hydrophilic surface, and the hydrophilic surface is relatively far away from the silicon wafer; dropping a colloidal solution on the hydrophilic surface of the antireflection optical film, wherein the colloidal solution includes a solution and multiple nano-balls and the nano-balls are adhered onto the hydrophilic surface; and performing an etching process on the hydrophilic surface of the antireflection optical film by taking the nano-balls as an etching mask so as to form a second rough surface, wherein the roughness of the second rough surface is different from the roughness of the first rough surface.