Investigation of image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy

Yoshinori Iketaki; Takeshi Watanabe; Nándor Bokor; Masaaki Fujii

doi:10.1117/12.698451

14 February 2007 Investigation of image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy

Yoshinori Iketaki, Takeshi Watanabe, Nándor Bokor, Masaaki Fujii

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Proceedings Volume 6443, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIV; 644307 (2007) https://doi.org/10.1117/12.698451
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

We quantitatively investigated image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy. To evaluate the properties, the point spread function (PSF) and contrast transfer function (CTF) were measured using a fluorescent scale together with a fluorescent bead. From the CTF, it has been found that visible light can resolve a 100 nm line-and-space pattern by microcopy, and provide a contrast of 10%. The CTF corresponds to a PSF with a FWHM of 130 nm. The value is two times finer than the diffraction limit size. An evaluation using a 100 nm Φ fluorescent bead consistently supports the result given by the CTF for 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 a spatial resolution overcoming the diffraction limit.

Citation Download Citation

Yoshinori Iketaki, Takeshi Watanabe, Nándor Bokor, and Masaaki Fujii "Investigation of image properties in super-resolution microscopy using two-color fluorescence dip spectroscopy", Proc. SPIE 6443, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIV, 644307 (14 February 2007); https://doi.org/10.1117/12.698451

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KEYWORDS

Luminescence

Contrast transfer function

Super resolution microscopy

Super resolution

Point spread functions

Diffraction

Fluorescence spectroscopy

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