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Instrumentation, Measurement, and Metrology

Theoretical investigation of the point-spread function given by super-resolving fluorescence microscopy using two-color fluorescence dip spectroscopy

[+] Author Affiliations
Yoshinori Iketaki

Olympus Co., Ltd., 2-3 Kuboyama-cho, Hachioji-shi 192-8512, Japan E-mail: y_iketaki@ot.olympus.co.jp

Takeshi Watanabe, Makoto Sakai, Shun-ichi Ishiuchi, Masaaki Fujii

Tokyo Institute of Technology, Chemical Resources Laboratory, 4259 Nagatsuta, Midori-ku, Yokohama 226-8503, Japan

Tsutomu Watanabe

University of Electro-Communications, Institute for Laser Science, 1-5-1 Chofugaoka, Chofu-shi 181-8585, Japan

Opt. Eng. 44(3), 033602 (Mar. 17, 2005). doi:10.1117/1.1871512
History: Received May 23, 2004; Revised Sep. 7, 2004; Accepted Sep. 8, 2004; Mar. 17, 2005; Online March 17, 2005
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The profile of the point spread function (PSF) in superresolution microscopy is studied theoretically. The fluorescence spot profile (i.e., the PSF) is determined by the focused beam patterns of the applied two-color lasers and the optical properties of the fluorescence-depletion process induced by the lasers (the pump and erase beams). In this study, the fluorescence-depletion process for the sample molecule is analyzed using a rate equation for a three-state model. Based on this result, we calculate the PSF for the case where the erase beam is modeled by a first-order Bessel function. In the case of an erase beam with a large photon flux, the obtained PSF has a Lorentzian-like shape, which seldom appears in traditional microscopy. In this work, we also investigated a possible relationship between the PSF and other parameters in the fluorescence-depletion process. © 2005 Society of Photo-Optical Instrumentation Engineers.

© 2005 Society of Photo-Optical Instrumentation Engineers

Citation

Yoshinori Iketaki ; Takeshi Watanabe ; Makoto Sakai ; Shun-ichi Ishiuchi ; Masaaki Fujii, et al.
"Theoretical investigation of the point-spread function given by super-resolving fluorescence microscopy using two-color fluorescence dip spectroscopy", Opt. Eng. 44(3), 033602 (Mar. 17, 2005). ; http://dx.doi.org/10.1117/1.1871512


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