A method of forming an enhanced super-resolution image is provided. The method includes: preparing a substrate comprising a glass substrate, a metal layer on the glass substrate, and a biolayer on the metal layer; placing a biological specimen on the substrate, the biological specimen being in contact with the biolayer and being attached to the metal layer through the biolayer, in which the biological specimen is labeled by a plurality of spontaneous blinking elements therein; irradiating a light beam to the metal layer via the glass substrate; receiving fluorescence signals emitted from the spontaneous blinking elements within a time period; fitting a plurality of functions respectively to each of the fluorescence signals; pinpointing peak positions of the functions; and reconstructing the peak positions to derive the enhanced super-resolution image of an underlying structure of the biological specimen in proximity to the metal layer.