The contrast transfer function (CTF) of super-resolution microscopy was quantitatively investigated using a fluorescent scale. The scale has minute fluorescent line patterns, finer than 100 nm, and is suitable for measuring CTF in super-resolution microscopy. The measured CTF shows that super-resolution microscopy can indeed improve the optical properties of fluorescent images and enable us to observe a structure with the spatial resolution overcoming the diffraction limit. From the CTF, it has been found that super-resolution microcopy can resolve a 100 nm line-and-space pattern and provides a contrast of 10%. The CTF corresponds to a PSF with a full-width at half-maximum (FWHM) of 130 nm. An evaluation using a 100 nmΦ fluorescent bead consistently supports the results given by the CTF for super-resolution microscopy.

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