Measurement of contrast transfer function in super-resolution microscopy using two-color fluorescence dip spectroscopy.
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 nmphi fluorescent bead consistently supports the results given by the CTF for super-resolution microscopy.
Iketaki
Yoshinori
Y
Olympus Corporation, 2-3 Kuboyama-cho, Hachioji, 192-8512, Japan. y_iketaki@ot.olympus.co.jp
Watanabe
Takeshi
T
Bokor
Nándor
N
Omatsu
Takashige
T
Hiraga
Takashi
T
Yamamoto
Kimihisa
K
Fujii
Masaaki
M
D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation* / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity