High-Resolution Confocal Microscopy by Saturated Excitation of Fluorescence

Katsumasa Fujita, Minoru Kobayashi, Shogo Kawano, Masahito Yamanaka, and Satoshi Kawata
Phys. Rev. Lett. 99, 228105 – Published 30 November 2007

Abstract

We demonstrate the use of saturated excitation in confocal fluorescence microscopy to improve the spatial resolution. In the proposed technique, we modulate the excitation intensity temporally and detect the harmonic modulation of the fluorescence signal which is caused by the saturated excitation in the center of the laser focus. Theoretical and experimental investigations show that the demodulated fluorescence signal is nonlinearly proportional to the excitation intensity and contributes to improve the spatial resolution in three dimensions beyond the diffraction limit of light.

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  • Received 15 August 2006

DOI:https://doi.org/10.1103/PhysRevLett.99.228105

©2007 American Physical Society

Authors & Affiliations

Katsumasa Fujita1,*, Minoru Kobayashi2, Shogo Kawano3, Masahito Yamanaka3, and Satoshi Kawata1,4

  • 1Department of Applied Physics, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871 Japan
  • 2Nanophoton Corp., A-509, CASI, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871 Japan
  • 3Department of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871 Japan
  • 4Nanophotonics Laboratory, RIKEN, Hirosawa, Wako, Saitama 351-0198 Japan

  • *fujita@ap.eng.osaka-u.acjp

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Vol. 99, Iss. 22 — 30 November 2007

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